Substituted carboxamides

ABSTRACT

This application relates to a compound of formula I (or a pharmaceutically acceptable salt thereof) as defined herein, pharmaceutical compositions thereof, and its use as an inhibitor of factor Xa, as well as a process for its preparation and intermediates therefor.

This application is the national stage of PCT/US01/42941, filed Nov. 14,2001, and claims the benefit of U.S. Provisional Application No.60/253,501, filed Nov. 28, 2000, which is incorporated by referenceherein.

This invention relates to anticoagulant substituted carboxamides whichdemonstrate activity as inhibitors of factor Xa and, accordingly, whichare useful antithrombotics in mammals. In particular it relates tosubstituted carboxamides having high anticoagulant activity, andantithrombotic activity. Thus, this invention relates to new substitutedcarboxamides which are inhibitors of factor Xa, pharmaceuticalcompositions containing the substituted carboxamides as activeingredients, and the use of the substituted carboxamides asanticoagulants for prophylaxis and treatment of thromboembolic disorderssuch as venous thrombosis, pulmonary embolism, arterial thrombosis, inparticular myocardial ischemia, myocardial infarction and cerebralthrombosis, general hypercoagulable states and local hypercoagulablestates, such as following angioplasty and coronary bypass operations,and generalized tissue injury as it relates to the inflammatory process.In addition, the substituted carboxamides are useful as anticoagulantsin in vitro applications.

The process of blood coagulation, thrombosis, is triggered by a complexproteolytic cascade leading to the formation of thrombin. Thrombinproteolytically removes activation peptides from the Aα-chains and theBβ-chains of fibrinogen, which is soluble in blood plasma, initiatinginsoluble fibrin formation. The formation of thrombin from prothrombinis catalyzed by factor Xa.

Anticoagulation currently is achieved by the administration of heparinsand coumarins. Parenteral pharmacological control of coagulation andthrombosis is based on inhibition of thrombin through the use ofheparins. Heparins act indirectly on thrombin by accelerating theinhibitory effect of endogenous antithrombin III (the main physiologicalinhibitor of thrombin). Because antithrombin III levels vary in plasmaand because clot-bound thrombin seems resistant to this indirectmechanism, heparins can be an ineffective treatment. Because coagulationassays are believed to be associated with efficacy and with safety,heparin levels must be monitored with coagulation assays (particularlythe activated partial thromboplastin time (APTT) assay). Coumarinsimpede the generation of thrombin by blocking the posttranslationalgamma-carboxylation in the synthesis of prothrombin and other proteinsof this type. Because of their mechanism of action, the effect ofcoumarins can only develop slowly, 6–24 hours after administration.Further, they are not selective anticoagulants. Coumarins also requiremonitoring with coagulation assays (particularly the prothrombin time(PT) assay).

Recently, interest has grown in small synthetic molecules whichdemonstrate potent direct inhibition of thrombin and factor Xa. See, B.Y. Zhu; R. M. Scarborough Current Opinion in Cardiovascular, Pulmonary &Renal Investigational Drugs, (1999), 1(1), 63–88, Recent Advances inInhibitors of Factor Xa in the Prothrombinase Complex.

Although the heparins and coumarins are effective anticoagulants, therestill exists a need for anticoagulants which act selectively on factorXa or thrombin, and which, independent of antithrombin III, exertinhibitory action shortly after administration, preferably by an oralroute, and do not interfere with lysis of blood clots, as required tomaintain hemostasis.

The present invention is directed to the discovery that the substitutedheterocyclic amides of the present invention, as defined below, arepotent inhibitors of factor Xa which may have high bioavailabilityfollowing oral administration.

According to the invention there is provided a compound of formula I

or a pharmaceutically acceptable salt thereof, wherein

R¹ is 2-pyridinyl (which may bear a methyl, methoxy, methylthio, fluoroor chloro substituent at the 5-position), or R¹ is 3-pyridinyl (whichmay bear a methyl, fluoro or chloro substituent at the 6-position), orR¹ is phenyl (which may bear one, two or three substituents at the 3-,4- or 5-position(s) independently selected from halo, cyano, carbamoyl,methyl, methoxy., difluoromethoxy, hydroxymethyl, formyl, vinyl, amino,hydroxy and 3, 4-methylenedioxy; and in addition the phenyl may bear a2-chloro or 2-fluoro substituent), or R¹ is 6-indolyl (which may bear achloro or methyl substituent at the 3-position), or R¹ is 6-indazolyl(which may bear a methyl substituent at the 3-position);

A³, A⁴, A⁵ and A⁶, together with the two carbons to which they areattached, complete a substituted benzene in which A³ is CR³, A⁴ is CR⁴,A⁵ is CR⁵, and A⁶ is CR⁶;

wherein

R³ is hydrogen, fluoro, chloro, methyl, methoxy, hydroxy or carboxy;

one of R⁴ and R⁵ is hydrogen, (1–4C)alkyl, halo, trifluoromethyl,trifluoromethoxy, cyano, hydroxymethyl, (1–3C)acyl, R^(f)O—, R^(f)O₂C—,R^(f)O₂C—CH₂—, R^(f)O₂C—CH₂—O—, methylthio or R^(g)NH—;

the other of R⁴ and R⁵ is hydrogen, halo or methyl; and

R⁶ is hydrogen, fluoro, chloro, methyl or methoxy;

in which R^(f) is hydrogen, (1–4C)alkyl or benzyl; R^(g) is hydrogen,(1–3C)acyl, trifluoroacetyl, methoxyacetyl, or R^(h)SO_(h)— (wherein his 1 or 2); and R^(h) is (1–4C)alkyl, trifluoromethyl, phenyl, amino,methylamino or dimethylamino; or

A³, A⁴, A⁵ and A⁶, together with the two carbons to which they areattached, complete a substituted heteroaromatic ring in which

(a) one of A³, A⁴, A⁵ and A⁶ is N, and each of the others is CR³, CR⁴,CR⁵ or CR⁶, respectively; wherein

each of R³, R⁴, R⁵ and R⁶ is independently hydrogen or methyl, or one ofR³, R⁴, R⁵ and R⁶ attached to a carbon which is not bonded to an N-atomis chloro and the others are hydrogen; or

(b) two adjacent residues of A³, A⁴, A⁵ and A⁶ together form S, and eachof the others is CR³, CR⁴, CR⁵ or CR⁶, respectively; wherein

each of R³, R⁴, R⁵ and R⁶ is hydrogen, or one or two of R³, R⁴, R⁵ andR⁶ is independently chloro, bromo or methyl and the others are hydrogen;or

(c) A³ and A⁴ together form a fused benz ring, and A⁵ and A⁶ togetherform —NH—;

L is carbonyl or methylene; M is N and Q (showing L and M at the pointsof attachment) is a residue of formula Q^(A),

in which

each of X¹, X², X³ and X⁴ is hydrogen; or

X¹ is OR^(Q) and each of X², X³ and X⁴ is hydrogen in which R^(Q) ishydrogen, (1–3C)alkyl, (3–6C)cycloalkyl, 2-hydroxyethyl, 2-methoxyethylor 2-methylthioethyl; or

one of X¹ and X² is trifluoromethyl; and each of the others of X¹, X²,X³ and X⁴ is hydrogen; or

one or two of X² and X³ is fluoro; and each of the others of X¹, X², X³and X⁴ is hydrogen; or

L is carbonyl or methylene; M is N; and Q is cyclohexan-1,4-diyl; or

L is carbonyl or methylene; M is CH and Q is piperidin-1,4-diyl which isbonded to L at the 4-position and is bonded to M at the 1-position; or

L is carbonyl; M is N and Q is piperidin-1,4-diyl which is bonded to Lat the 1-position and is bonded to M at the 4-position; and

R is hydrogen, (1–3C)alkyl, (3–5C)cycloalkyl, (1–3C)acyl,acetyloxyacetyl, aminoacetyl, hydroxyacetyl, {(1–4C)alkoxy}carbonyl,{(1–4C) alkoxy}carbonylmethyl, R^(a)R^(b)N—CO— or R^(j)SO_(j)—, whereineach of R^(a) and R^(b) is independently hydrogen or (1–3C)alkyl, orR^(a)R^(b)N— is 1-azetidinyl, 1-pyrrolidinyl, 1-piperidinyl,4-morpholinyl, or 4-thiomorpholinyl; j is 1 or 2; and R^(j) is(1–4C)alkyl, trifluoromethyl, amino, methylamino or dimethylamino.

As used herein, the expression a compound of formula I or the expressiona compound of the invention includes the compound and any conventionalprodrug thereof, as well as a pharmaceutically acceptable salt of saidcompound or prodrug.

In this specification, the following definitions are used, unlessotherwise described: Halo is fluoro, chloro, bromo or iodo. Alkyl,alkoxy, etc. denote both straight and branched groups; but reference toan individual radical such as “propyl” embraces only the straight chain(“normal”) radical, a branched chain isomer such as “isopropyl” beingspecifically denoted.

Particular values are listed below for radicals, substituents, andranges, for illustration only, and they do not exclude other definedvalues or other values within defined ranges for the radicals andsubstituents. Thus, a particular value for halo is fluoro, chloro orbromo; for (1–3C)alkyl is methyl, ethyl, propyl or isopropyl; for(1–4C)alkyl is methyl, ethyl, propyl, isopropyl, butyl, isobutyl ort-butyl; for (3–5C)cycloalkyl is cyclopropyl, cyclobutyl or cyclopentyl;for (3–6C)cycloalkyl is cyclopropyl, cyclopentyl or cyclohexyl; for(1–4C)alkoxy is methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxyor t-butoxy; and for (1–3C)acyl is formyl, acetyl or propionyl.

A particular compound of formula I is one wherein:

R¹ is 2-pyridinyl (which may bear a methyl, methoxy, methylthio, fluoroor chloro substituent at the 5-position), or R¹ is 3-pyridinyl (whichmay bear a methyl, fluoro or chloro substituent at the 6-position), orR¹ is phenyl (which may bear a substituent at the 3- or 4-position(s)independently selected from halo, cyano, carbamoyl, methyl, methoxy,difluoromethoxy, hydroxymethyl, formyl, vinyl, amino, hydroxy and3,4-methylenedioxy), or R¹ is 6-indolyl (which may bear a chloro ormethyl substituent at the. 3-position);

A³ is CR³, A⁴ is CR⁴, A⁵ is CR⁵, and A⁶ is CR⁶; wherein

R³ is hydrogen;

one of R⁴ and R⁵ is hydrogen, (1–4C)alkyl, halo, trifluoromethyl,trifluoromethoxy, cyano, hydroxymethyl, (1–3C)acyl, R^(f)O—, R^(f)O₂C—,R^(f)O₂C—CH₂—, R^(f)O₂C—CH₂—O—, methylthio or R^(g)NH— in which R^(g) ishydrogen, (1–3C)acyl, or R^(h)SO₂—; and R^(h) is (1–4C)alkyl,trifluoromethyl, amino, methylamino or dimethylamino;

the other of R⁴ and R⁵ is hydrogen, halo or methyl; and

R⁶ is hydrogen; or

A³ is N, and each of A⁴, A⁵ and A⁶ is CR⁴, CR⁵ and CR⁶, respectively, inwhich each of R⁴ and R⁶ is hydrogen and R⁵ is hydrogen, chloro ormethyl; or

A⁶ is N, and each of A³, A⁴ and A⁵ is CR³, CR⁴ and CR⁵, respectively, inwhich each of R³ and R⁴ is hydrogen and R⁵ is hydrogen or methyl; and

R is hydrogen, methyl, ethyl, acetyl, acetoxyacetyl, hydroxyacetyl,methylsulfonyl or dimethylaminosulfonyl.

A more particular compound, or salt thereof, as described above in onewherein:

R¹ is 2-pyridinyl which bears a methyl or chloro substituent at the5-position,

A³ is CR³, A⁴ is CR⁴, A⁵ is CR⁵, and A⁶ is CR⁶ wherein each of R³, R⁴and R⁶ is hydrogen and R⁵ is fluoro, chloro or methyl; or

A³ is N, and each of A⁴, A⁵ and A⁶ is CH; or

A⁶ is N, and each of A³, A⁴ and A⁵ is CH; and

R is hydrogen or methyl.

One particular compound, or salt thereof, according to any of the abovedescriptions is one wherein Q is Q^(A); and more particularly wherein Qis Q^(A) in which:

each of X¹, X², X³ and X⁴ is hydrogen; or

X¹ is OR^(Q) and each of X², X³ and X⁴ is hydrogen in which R^(Q) ishydrogen, (1–3C)alkyl, (3–6C)cycloalkyl, 2-methoxyethyl or2-methylthioethyl; or

X¹ is trifluoromethyl and each of X², X³ and X⁴ is hydrogen; or

X² is trifluoromethyl and each of X¹, X³ and X⁴ is hydrogen; or

X² is fluoro and each of X¹, X³ and X⁴ is hydrogen; or

each of X² and X³ is fluoro and each of X¹ and X⁴ is hydrogen.

One such compound of formula I is one wherein each of X¹, X², X³ and X⁴is hydrogen.

Another such compound of formula I is one wherein X¹ is OR^(Q) and eachof X², X³ and X⁴ is hydrogen in which R^(Q) is hydrogen, ethyl,isopropyl, 2-hydroxyethyl, 2-methoxyethyl or 2-methylthioethyl.

Another such compound of formula I is one wherein X¹ is trifluoromethyland each of X², X³ and X⁴ is hydrogen.

Another such compound of formula I is one wherein X² is trifluoromethyland each of X¹, X³ and X⁴ is hydrogen.

Another such compound of formula I is one wherein each of X² and X³ isfluoro and each of X¹ and X⁴ is hydrogen.

Another particular compound, or salt thereof, according to any of theabove descriptions is one wherein Q is cyclohexan-1,4-diyl.

A further particular compound, or salt thereof, according to any of theabove descriptions is one wherein Q is piperidin-1,4-diyl which isbonded to L at the 4-position and is bonded to M at the 1-position.

For any of the above descriptions a more particular compound or saltthereof of formula I is one where L is carbonyl.

Another more particular compound, or salt thereof, for any of the abovedescriptions is one wherein L is methylene.

An additional particular compound, or salt thereof, according to any ofthe above descriptions is one wherein Q is piperidin-1,4-diyl which isbonded to L at the 1-position and is bonded to M at the 4-position.

A specific compound, or salt thereof, wherein Q is Q^(A) is any one ofthose provided in the Examples, especially that of Example 4, 8, 9, 10,18, 19 or 21.

A pharmaceutically acceptable salt of a compound of the instantinvention is one which is the acid addition salt of a basic compound offormula I with an inorganic or organic acid which affords aphysiologically acceptable anion or which is the salt formed by anacidic compound of formula I with a base which affords a physiologicallyacceptable cation and provides a particular aspect of the invention.Examples of such acids and bases are provided hereinbelow.

As an additional aspect of the invention there is provided apharmaceutical formulation comprising in association with apharmaceutically acceptable carrier, diluent or excipient, a compound offormula I (or a pharmaceutically acceptable salt thereof) as provided inany of the descriptions herein.

In addition, there is provided the use of a compound of formula I (orprodrug or salt) as described herein as an active ingredient in themanufacture of a medicament for use in producing an anticoagulant orantithrombotic effect.

The present invention also provides a method of inhibiting coagulationin a mammal comprising administering to a mammal in need of treatment, acoagulation inhibiting dose of a compound of formula I having any of thedefinitions herein.

The present invention further provides a method of inhibiting factor Xacomprising administering to a mammal in need of treatment, a factor Xainhibiting dose of compound of formula I having any of the definitionsherein.

Further, the present invention provides a method of treating athromboembolic disorder comprising administering to a mammal in need oftreatment, an effective dose of a compound of formula I having any ofthe definitions herein.

Also, there is provided a compound of formula I (or prodrug or salt)having any of the definitions herein for use as an antithrombotic agent.

In addition, there is provided the use of a compound of formula I havingany of the definitions herein for the manufacture of a medicament fortreatment of a thromboembolic disorder.

As an additional feature of the invention there is provided apharmaceutical formulation comprising in association with apharmaceutically acceptable carrier, diluent or excipient, a prodrug ofa compound of formula I (or of a pharmaceutically acceptable saltthereof) as provided in any of the descriptions herein.

It will be appreciated that certain compounds of formula I (or salts orprodrugs, etc.) may exist in, and be isolated in, isomeric forms,including tautomeric forms, cis- or trans-isomers, as well as opticallyactive, racemic, or diastereomeric forms. It is to be understood thatthe present invention encompasses a compound of formula I in any of thetautomeric forms or as an a mixture thereof; or as a mixture ofdiastereomers, as well as in the form of an individual diastereomer, andthat the present invention encompasses a compound of formula I as amixture of enantiomers, as well as in the form of an individualenantiomer, any of which mixtures or form possesses inhibitoryproperties against factor Xa, it being well known in the art how toprepare or isolate particular forms and how to determine inhibitoryproperties against factor Xa by standard tests including those describedbelow.

In addition, a compound of formula I (or salt or prodrug, etc.) mayexhibit polymorphism or may form a solvate with water or an organicsolvent. The present invention also encompasses any such polymorphicform, any solvate or any mixture thereof.

A prodrug of a compound of formula I may be one formed in a conventionalmanner with a functional group of the compound, such as with an amino,hydroxy or carboxy group.

A compound of formula I may be prepared by processes which includeprocesses known in the chemical art for the production of structurallyanalogous compounds or by a novel process described herein. A novelprocess described herein provides another aspect of the invention. Aprocess for the preparation of a compound of formula I (or apharmaceutically acceptable salt thereof) and novel intermediates forthe manufacture of a compound of formula I provide further features ofthe invention and are illustrated by the following procedures in whichthe meanings of the generic radicals are as defined above, unlessotherwise specified. It will be recognized that it may be preferred ornecessary to prepare a compound of formula I in which a functional groupis protected using a conventional protecting group, then to remove theprotecting group to provide the compound of formula I.

Thus, there is provided a process for preparing a compound of formula I,or a pharmaceutically acceptable salt thereof, as provided in any of theabove descriptions, which comprises:

(A) for a compound of formula I in which Q is Q^(A), substituting thegroup Y^(a) of a compound of formula II,

in which Y^(a) is a leaving group for nucleophilic aromaticsubstitution, using an amine of formula III,

(B) for a compound of formula I in which L is carbonyl, acylating anamine of formula IV

using a corresponding acid of formula V,

or an activated derivative thereof;

(C) for a compound of formula I in which R is not hydrogen, substitutingthe nitrogen of a corresponding compound in which R is hydrogen using aconventional procedure;

(D) for a compound of formula I in which L is methylene, substitutingthe group Y^(a) of a compound of formula VI

in which Y^(a) is a leaving group for nucleophilic aromatic substitutionwith an amine of formula VII; or

alkylating an amine of formula IV directly, using a compound of formulaVIII,

in which Y^(b) is a leaving group for nucleophilic substitution, orindirectly by reductive alkylation using an aldehyde of formula IX;

(E) acylating an amine of formula H₂N—R¹, or a deprotonated derivativethereof, using an acid of formula X,

or an activated derivative thereof; or

(F) for a compound of formula I in which Q is cyclohexan-1,4-diyl or Qis piperidin-1,4-diyl which is bonded to L at the 1-position,reductively alkylating an amine of formula III using a compound offormula XIa or XIb, respectively;

(G) for a compound of formula I in which Q is piperidin-1,4-diyl whichis bonded to L at the 4-position, reductively alkylating an amine offormula XII

using a compound of formula XIII;

whereafter, for any of the above procedures, when a functional group ofa starting material is protected using a protecting group, removing theprotecting group;

whereafter, for any of the above procedures, when a pharmaceuticallyacceptable salt of a compound of formula I is required, it is obtainedby reacting the basic form of a basic compound of formula I with an acidaffording a physiologically acceptable counterion or the acidic form ofan acidic compound of formula I with a base affording a physiologicallyacceptable counterion or by any other conventional procedure;

and wherein, unless otherwise specified, R¹, A³–A⁶, L, M, Q, and R haveany of the values defined in any of-the above definitions.

As used herein, a leaving group “Y^(a)” is a moiety which is displacedin an aromatic (or heteroaromatic) nucleophilic substitution reaction,for example a halo group (such as fluoro or chloro), an alkoxy group(such as methoxy), a sulfonate ester group (such as methylsulfonyloxy,p-toluylsulfonyloxy or trifluoromethylsulfonyloxy), or the reactivespecies derived from treating an alcohol with triphenylphospine, diethylazodicarboxylate and triethyl amine (in a Mitsunobu reaction). Thesubstitution may be carried out by heating a mixture of the reagents ina polar solvent, for example in dimethyl sulfoxide in a sealed tube orvial as described at Example 5, Example 18a, and Example 22.

For a carboxylic acid, a typical activated derivative includes an ester(particularly a lower alkyl ester such as the methyl or ethyl ester), anacid halide (particularly the acid chloride), and an activated ester oranhydride (including the 4-nitrophenyl ester and an activated ester oranhydride derived from a coupling reagent), as well as (when the productis a urea) the isocyanate. Typical procedures include those describedfor preparation of intermediate compounds at Intermediate A-2 andExample 23-A, as well as the O-protected product of Example 17.

For a compound of formula I in which R is (1-C)alkyl or{(1–4C)alkoxy}carbonylmethyl, a conventional procedure for substitutingthe nitrogen of a compound in which R is hydrogen comprises alkylatingthe nitrogen, for example by reductively alkylating the nitrogen withthe requisite aldehyde or ketone or by alkylating the nitrogen with areagent of formula R-Y^(b), in which Y^(b) is a leaving group fornucleophilic substitution.

For a compound of formula I in which R is (1–3C)acyl, acetyloxyacetyl,aminoacetyl, hydroxyacetyl, {(1–4C)alkoxy}-carbonyl or R^(a)R^(b)N—CO—,a conventional procedure for substituting the nitrogen of a compound inwhich R is hydrogen comprises acylating the nitrogen using the requisitecarboxylic acid or activated derivative thereof (in which a functionalgroup may be protected, followed by removal of the protecting group).

For a compound of formula I in which R is R^(j)SO_(j)—, a conventionalprocedure for substituting the nitrogen of a compound in which R ishydrogen comprises treating the amine with the requisite sulfinyl orsulfonyl halide, for example using the chloride of formulaR^(j)SO_(j)—Cl.

As used herein, a leaving group “Y^(b)” is a moiety which is displacedin a nucleophilic substitution reaction, for example a halo group (suchas chloro, bromo or iodo), a sulfonate ester group (such asmethylsulfonyloxy, p-toluylsulfonyloxy or trifluoromethylsulfonyloxy),or the reactive species derived from treating an alcohol withtriphenylphospine, diethyl azodicarboxylate and triethyl amine (in aMitsunobu reaction).

A novel intermediate or starting material compound provides a furtheraspect of the invention. The various starting materials may be made byprocesses which include processes known in the chemical art for theproduction of structurally analogous compounds or by a novel processdescribed herein or one analogous thereto.

Thus, one particular intermediate is a compound of formula II,

in which Y^(a) is a leaving group for nucleophilic aromatic substitutionand R¹, A³–A⁶, L and Q^(A) have any of the values defined in any of theabove definitions, or a derivative thereof in which a functional groupother than Y^(a) is protected using a protecting group. It is preferredthat the intermediate of formula II be one in which L is carbonyl.

Another intermediate is an acid of formula V,

or an activated derivative thereof, or a salt of the acid or activatedderivative, in which Q and R have any of the values defined in any ofthe above definitions, or a derivative thereof in which a functionalgroup other than the carboxy group, or activated derivative thereof, isprotected using a protecting group.

A further intermediate is an amine of formula VII, or a salt thereof; or

a compound of formula VIII, or a salt thereof,

in which Y^(b) is a leaving group for nucleophilic substitution, oran aldehyde of formula IX, or a salt thereof;

in which M, Q and R have any of the values defined in any the abovedefinitions, or a derivative thereof in which a functional group otherthan the primary amino group of the compound of formula VII, the leavinggroup of the compound of formula VIII, or the aldehyde group of thecompound of formula IX, respectively, is protected using a protectinggroup.

Another intermediate is an acid of formula X,

or an activated derivative thereof, or a salt of the acid or activatedderivative, in which A³–A⁶, L, M, Q and R have any of the values definedin any of the above definitions, or a derivative thereof in which afunctional group other than the carboxy group, or activated derivativethereof, is protected using a protecting group; and more particularlythe acid of formula X in which L is carbonyl.

For an acid of formula X in which L is carbonyl, a particular activatedderivative is a compound of formula Xa

and for an acid of formula X in which L is methylene, a particularactivated derivative is a compound of formula Xb

or a salt of the activated derivative, in which A³–A⁶, M, Q and R haveany of the values defined in any of the above descriptions, or aderivative thereof in which a functional group other than the activatedderivative of the carboxy group is protected using a protecting group.

A further particular intermediate is a compound of formula XIa,

or a salt thereof, wherein R¹, A³–A⁶ and L have any of the valuesdefined in any of the above descriptions; and, more particularly, thecompound, or salt thereof, in which L is carbonyl.

As a further aspect of the invention, there is provided the use of acompound (or activated and/or protected derivative thereof or salt ofthe compound or derivative) of formula II, V, VII, VIII, IX, X or XIa asa starting material in the preparation of an inhibitor of factor Xa.

As mentioned above, a compound corresponding to a compound of formula Ibut in which a functional group is protected may serve as anintermediate for a compound of formula I. Accordingly, such a protectedintermediate for a novel compound of formula I provides a further aspectof the invention. Thus, as one particular aspect of the invention, thereis provided a compound corresponding to a novel compound of formula I asdefined above in which there is a hydroxy, but in which thecorresponding substituent is —OP^(p) in place of hydroxy, wherein P^(p)is a phenol protecting group other than methyl. Phenol protecting groupsare well known in the art, for example as described in T. W. Greene andP. G. M. Wuts, “Protecting Groups in Organic Synthesis” (1991). Further,P^(p) may denote a functionalized resin, for example as disclosed in H.V. Meyers, et al., Molecular Diversity, (1995), 1, 13–20.

As mentioned above, the invention includes a pharmaceutically acceptablesalt of the factor Xa inhibiting compound defined by the above formulaI. A basic compound of this invention possesses one or more functionalgroups sufficiently basic to react with any of a number of inorganic andorganic acids affording a physiologically acceptable counterion to forma pharmaceutically acceptable salt. Acids commonly employed to formpharmaceutically acceptable acid addition salts are inorganic acids suchas hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid,phosphoric acid, and the like, and organic acids such asp-toluenesulfonic acid, methanesulfonic acid, oxalic acid,p-bromobenzenesulfonic acid, carbonic acid, succinic acid, citric acid,benzoic acid, acetic acid, and the like. Examples of suchpharmaceutically acceptable salts thus are the sulfate, pyrosulfate,bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate,dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide,iodide, acetate, propionate, decanoate, caprylate, acrylate, formate,isobutyrate, caproate, heptanoate, propiolate, oxalate, malonate,succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate,hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate,dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate,xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate,citrate, lactate, gamma-hydroxybutyrate, glycollate, tartrate,methanesulfonate, propanesulfonate, naphthalene-1-sulfonate,naphthalene-2-sulfonate, mandelate, and the like. Preferredpharmaceutically acceptable acid addition salts include those formedwith mineral acids such as hydrochloric acid, hydrobromic acid andsulfuric acid.

If not commercially available, a necessary starting material for thepreparation of a compound of formula I may be prepared by a procedurewhich is selected from standard techniques of organic chemistry,including aromatic and heteroaromatic substitution and transformation,from techniques which are analogous to the syntheses of known,structurally similar compounds, and techniques which are analogous tothe above described procedures or procedures described in the Examples.It will be clear to one skilled in the art that a variety of sequencesis available for the preparation of the starting materials. Startingmaterials which are novel provide another aspect of the invention.

Selective methods of substitution, protection and deprotection are wellknown in the art for preparation of a compound such as one of formulaII.

Generally, a basic compound of the invention is isolated best in theform of an acid addition salt. A salt of a compound of formula I formedwith an acid such as one of those mentioned above is useful as apharmaceutically acceptable salt for administration of theantithrombotic agent and for preparation of a formulation of the agent.Other acid addition salts may be prepared and used in the isolation andpurification of the compounds.

As noted above, the optically active isomers and diastereomers of thecompounds of formula I are also considered part of this invention. Suchoptically active isomers may be prepared from their respective opticallyactive precursors by the procedures described above, or by resolving theracemic mixtures. This resolution can be carried out by derivatizationwith a chiral reagent followed by chromatography or by repeatedcrystallization. Removal of the chiral auxiliary by standard methodsaffords substantially optically pure isomers of the compounds of thepresent invention or their precursors. Further details regardingresolutions can be obtained in Jacques, et al., Enantiomers, Racemates,and Resolutions, John Wiley & Sons, 1981.

The compounds of the invention are believed to selectively inhibitfactor Xa over other proteinases and nonenzyme proteins involved inblood coagulation without appreciable interference with the body'snatural clot lysing ability (the compounds have a low inhibitory effecton fibrinolysis). Further, such selectivity is believed to permit usewith thrombolytic agents without substantial interference withthrombolysis and fibrinolysis.

The invention in one of its aspects provides a method of inhibitingfactor Xa in a mammal comprising administering to a mammal in need oftreatment an effective (factor Xa inhibiting) dose of a compound offormula I.

In another of its aspects, the invention provides a method of treating athromboembolic disorder comprising administering to a mammal in need oftreatment an effective (thromboembolic disorder therapeutic and/orprophylactic amount) dose of a compound of formula I.

The invention in another of its aspects provides a method of inhibitingcoagulation in a mammal comprising administering to a mammal in need oftreatment an effective (coagulation inhibiting) dose of a compound offormula I.

The factor Xa inhibition, coagulation inhibition and thromboembolicdisorder treatment contemplated by the present method includes bothmedical therapeutic and/or prophylactic treatment as appropriate.

In a further embodiment, the invention relates to treatment, in a humanor animal, of a condition where inhibition of factor Xa is required. Thecompounds of the invention are expected to be useful in mammals,including man, in treatment or prophylaxis of thrombosis andhypercoagulability in blood and tissues. Disorders in which thecompounds have a potential utility are in treatment or prophylaxis ofthrombosis and hypercoagulability in blood and tissues. Disorders inwhich the compounds have a potential utility, in treatment and/orprophylaxis, include venous thrombosis and pulmonary embolism, arterialthrombosis, such as in myocardial ischemia, myocardial infarction,unstable angina, thrombosis-based stroke and peripheral arterialthrombosis. Further, the compounds have expected utility in thetreatment or prophylaxis of atherosclerotic disorders (diseases) such ascoronary arterial disease, cerebral arterial disease and peripheralarterial disease. Further, the compounds are expected to be usefultogether with thrombolytics in myocardial infarction. Further, thecompounds have expected utility in prophylaxis for reocclusion afterthrombolysis, percutaneous transluminal angioplasty (PTCA) and coronarybypass operations. Further, the compounds have expected utility inprevention of rethrombosis after microsurgery. Further, the compoundsare expected to be useful in anticoagulant treatment in connection withartificial organs, including joint replacement, and cardiac valves.Further, the compounds have expected utility in anticoagulant treatmentin hemodialysis and disseminated intravascular coagulation. Further, thecompounds may be useful in reducing the increased thrombin generationwhich occurs in the airways of patients with asthma; see, E. C. Gabazza,et al., Lung, (1999), 177(4), 253–262. A further expected utility is inrinsing or coating of catheters and mechanical devices used in patientsin vivo, and as an anticoagulant for preservation of blood, plasma andother blood products in vitro. Still further, the compounds haveexpected utility in other diseases where blood coagulation could be afundamental contributing process or a source of secondary pathology,such as cancer, including metastasis, inflammatory diseases, includingarthritis, and diabetes. The anti-coagulant compound is administeredorally or parenterally, e.g. by intravenous infusion (iv), intramuscularinjection (im) or subcutaneously (sc).

The specific dose of a compound administered according to this inventionto obtain therapeutic and/or prophylactic effects will, of course, bedetermined by the particular circumstances surrounding the case,including, for example, the compound administered, the rate ofadministration, the route of administration, and the condition beingtreated.

A typical daily dose for each of the above utilities is between about0.01 mg/kg and about 1000 mg/kg. The dose regimen may vary e.g. forprophylactic use a single daily dose may be administered or multipledoses such as 3 or 5 times daily may be appropriate. In critical caresituations a compound of the invention is administered by iv infusion ata rate between about 0.01 mg/kg/h and about 20 mg/kg/h and preferablybetween about 0.1 mg/kg/h and about 5 mg/kg/h.

The method of this invention also is practiced in conjunction with aclot lysing agent e.g. tissue plasminogen activator (t-PA), modifiedt-PA, streptokinase or urokinase. In cases when clot formation hasoccurred and an artery or vein is blocked, either partially or totally,a clot lysing agent is usually employed. A compound of the invention canbe administered prior to or along with the lysing agent or subsequent toits use, and preferably further is administered along with aspirin toprevent the reoccurrence of clot formation.

The method of this invention is also practiced in conjunction with aplatelet glycoprotein receptor (IIb/IIIa) antagonist, that inhibitsplatelet aggregation. A compound of the invention can be administeredprior to or along with the IIb/IIIa antagonist or subsequent to its useto prevent the occurrence or reoccurrence of clot formation. The methodof this invention is also practiced in conjunction with aspirin. Acompound of the invention can be administered prior to or along withaspirin or subsequent to its use to prevent the occurrence orreoccurrence of clot formation. As stated above, preferably a compoundof the present invention is administered in conjunction with a clotlysing agent and aspirin.

This invention also provides a pharmaceutical composition for use in theabove described therapeutic method. A pharmaceutical composition of theinvention comprises an effective factor Xa inhibiting amount of acompound of formula I in association with a pharmaceutically acceptablecarrier, excipient or diluent.

The active ingredient in such formulations comprises from 0.1 percent to99.9 percent by weight of the formulation. By “pharmaceuticallyacceptable” it is meant the carrier, diluent or excipient must becompatible with the other ingredients of the formulation and notdeleterious to the recipient thereof.

For oral administration the antithrombotic compound is formulated ingelatin capsules or tablets which may contain excipients such asbinders, lubricants, disintegration agents and the like. For parenteraladministration the antithrombotic is formulated in a pharmaceuticallyacceptable diluent e.g. physiological saline (0.9 percent), 5 percentdextrose, Ringer's solution and the like.

The compound of the present invention can be formulated in unit dosageformulations comprising a dose between about 0.1 mg and about 1000 mg.Preferably, the compound is in the form of a pharmaceutically acceptablesalt such as for example the sulfate salt, acetate salt or a phosphatesalt. An example of a unit dosage formulation comprises 5 mg of acompound of the present invention as a pharmaceutically acceptable saltin a 10 mL sterile glass ampoule. Another example of a unit dosageformulation comprises about 10 mg of a compound of the present inventionas a pharmaceutically acceptable salt in 20 mL of isotonic salinecontained in a sterile ampoule.

The compounds can be administered by a variety of routes including oral,rectal, transdermal, subcutaneous, intravenous, intramuscular, andintranasal. The compounds of the present invention are preferablyformulated prior to administration.

The present pharmaceutical compositions are prepared by known proceduresusing well known and readily available ingredients. The compositions ofthis invention may be formulated so as to provide quick, sustained, ordelayed release of the active ingredient after administration to thepatient by employing procedures well known in the art. In making thecompositions of the present invention, the active ingredient willusually be admixed with a carrier, or diluted by a carrier, or enclosedwithin a carrier which may be in the form of a capsule, sachet, paper orother container. When the carrier serves as a diluent, it may be asolid, semi-solid or liquid material which acts as a vehicle, excipientor medium for the active ingredient. Thus, the compositions can be inthe form of tablets, pills, powders, lozenges, sachets, cachets,elixirs, suspensions, emulsions, solutions, syrups, aerosols, (as asolid or in a liquid medium), soft and hard gelatin capsules,suppositories, sterile injectable solutions, sterile packaged powders,and the like.

The following formulation examples are illustrative only and are notintended to limit the scope of the invention in any way. “Activeingredient,” of course, means a compound according to formula I or apharmaceutically acceptable salt or solvate thereof.

Formulation 1: Hard gelatin capsules are prepared using the followingingredients:

Quantity (mg/capsule) Active ingredient 250 Starch, dried 200 Magnesiumstearate 10 Total 460 mg

Formulation 2: A tablet is prepared using the ingredients below:

Quantity (mg/tablet) Active ingredient 250 Cellulose, microcrystalline400 Silicon dioxide, fumed 10 Stearic acid 5 Total 665 mgThe components are blended and compressed to form tablets each weighing665 mg.

Formulation 3: An aerosol solution is prepared containing the followingcomponents:

Weight Active ingredient 0.25 Ethanol 29.75 Propellant 22(Chlorodifluoromethane) 70.00 Total 100.00The active compound is mixed with ethanol and the mixture added to aportion of the propellant 22, cooled to −30° C. and transferred to afilling device. The required amount is then fed to a stainless steelcontainer and diluted with the remainder of the propellant. The valveunits are then fitted to the container.

Formulation 4: Tablets, each containing 60 mg of active ingredient, aremade as follows:

Active ingredient  60 mg Starch  45 mg Microcrystalline cellulose  35 mgPolyvinylpyrrolidone (as 10% solution in  4 mg water) Sodiumcarboxymethyl starch  4.5 mg Magnesium stearate  0.5 mg Talc  1 mg Total150 mgThe active ingredient, starch and cellulose are passed through a No. 45mesh U.S. sieve and mixed thoroughly. The aqueous solution containingpolyvinylpyrrolidone is mixed with the resultant powder, and the mixturethen is passed through a No. 14 mesh U.S. sieve. The granules soproduced are dried at 50° C. and passed through a No. 18 mesh U.S.Sieve. The sodium carboxymethyl starch, magnesium stearate and talc,previously passed through a No. 60 mesh U.S. sieve, are then added tothe granules which, after mixing, are compressed on a tablet machine toyield tablets each weighing 150 mg.

Formulation 5: Capsules, each containing 80 mg of active ingredient, aremade as follows:

Active ingredient  80 mg Starch  59 mg Microcrystalline cellulose  59 mgMagnesium stearate  2 mg Total 200 mgThe active ingredient, cellulose, starch, and magnesium stearate areblended, passed through a No. 45 mesh U.S. sieve, and filled into hardgelatin capsules in 200 mg quantities.

Formulation 6: Suppositories, each containing 225 mg of activeingredient, are made as follows:

Active ingredient   225 mg Saturated fatty acid glycerides 2,000 mgTotal 2,225 mgThe active ingredient is passed through a No. 60 mesh U.S. sieve andsuspended in the saturated fatty acid glycerides previously melted usingthe minimum heat necessary. The mixture is then poured into asuppository mold of nominal 2 g capacity and allowed to cool.

Formulation 7: Suspensions, each containing 50 mg of active ingredientper 5 mL dose, are made as follows:

Active ingredient   50 mg Sodium carboxymethyl cellulose   50 mg Syrup1.25 mL Benzoic acid solution 0.10 mL Flavor q.v. Color q.v. Purifiedwater to total   5 mLThe active ingredient is passed through a No. 45 mesh U.S. sieve andmixed with the sodium carboxymethyl cellulose and syrup to form a smoothpaste. The benzoic acid solution, flavor and color are diluted with aportion of the water and added, with stirring. Sufficient water is thenadded to produce the required volume.

The ability of a compound of the present invention to be an effectiveand orally active factor Xa inhibitor may be evaluated-in one or more ofthe following assays or in other standard assays known to those in theart.

The inhibition by a compound of the inhibition of a serine protease ofthe human blood coagulation system or of the fibrinolytic system, aswell as of trypsin, is determined in vitro for the particular enzyme bymeasuring its inhibitor binding affinity in an assay in which the enzymehydrolyzes a particular chromogenic substrate, for example as describedin Smith, G. F.; Gifford-Moore, D.; Craft, T. J.; Chirgadze, N.;Ruterbories, K. J.; Lindstrom, T. D.; Satterwhite, J. H. Efegatran: ANew Cardiovascular Anticoagulant. New Anticoagulants for theCardiovascular Patient; Pifarre, R., Ed.; Hanley & Belfus, Inc.:Philadelphia, 1997; pp. 265–300. The inhibitor binding affinity ismeasured as apparent association constant Kass which is the hypotheticalequilibrium constant for the reaction between enzyme and the testinhibitor compound (I).

$\begin{matrix}\left. {{Enzyme} + I}\rightleftarrows{{Enzyme} - I} \right. \\{{Kass} = \frac{\left\lbrack {{Enzyme} - I} \right\rbrack}{\left( {\lbrack{Enzyme}\rbrack \times \lbrack I\rbrack} \right)}}\end{matrix}$

Conveniently, enzyme inhibition kinetics are performed in a high-volumeprotocol using automated dilutions of inhibitors (n=3 for each of fourto eight inhibitor concentrations) into 96-well polystyrene plates andreaction rates are determined from the rate of hydrolysis of appropriatep-nitroanilide substrates at 405 nm using a Thermomax plate reader fromMolecular Devices (San Francisco, Calif.). The same general protocol isfollowed for all enzymes studied: In each well is placed 50 μL buffer(0.06 M Tris, 0.3 M NaCl, pH 7.4), followed by 25 μL of inhibitorsolution (in 100% methanol) and 25 μL enzyme solution (e.g., humanfactor Xa, 32 nM in 0.03 M Tris, 0.15 M NaCl, 1 mg/mL HAS); finally,within two minutes, 150 μL aqueous solution of chromogenic substrate(e.g., 0.3 mM BzIle-Glu-Gly-Arg-pNA) is added to start the enzymaticreaction. Final factor Xa concentration is 3.2 nM. The rates ofchromogenic substrate hydrolysis reactions provide a linear relationshipwith the enzymes studied such that free enzyme can be quantitated inreaction mixtures. Data is analyzed directly as rates by the Softmaxprogram to produce [free enzyme] calculations for tight-binding Kassdeterminations. For apparent Kass determinations, human factor Xa isused to hydrolyze BzIle-Glu-Gly-Arg-pNA; 5.9 nM human thrombin is usedto hydrolyze 0.2 mM BzPhe-Val-Arg-pNA; 3.4 nM human plasmin is used with0.5 mM HD-Val-Leu-Lys-pNA; 1.2 nM human nt-PA is used with 0.8 mMHD-Ile-Pro-Arg-pNA; and 0.4 nM urokinase is used with 0.4 mMpyro-Glu-Gly-Arg-pNA.

Kass is calculated for a range of concentrations of test compounds whichproduce hydrolysis inhibition of between 20% and 80% of control and themean value reported in units of liter per mole. In general, a factor Xainhibiting compound of formula I of the instant invention, asexemplified herein, exhibits a Kass of greater than 10⁶ L/mole and acompound of formula I in which Q is Q^(A) exhibits a Kass of 10×10⁶L/mole or much greater.

The factor Xa inhibitor preferably should spare fibrinolysis induced byurokinase, tissue plasminogen activator (t-PA) and streptokinase. Thiswould be important to the therapeutic use of such an agent as an adjunctto streptokinase, tp-PA or urokinase thrombolytic therapy and to the useof such an agent as an endogenous fibrinolysis-sparing (with respect tot-PA and urokinase) antithrombotic agent. In addition to the lack ofinterference with the amidase activity of the fibrinolytic proteases,such fibrinolytic system sparing can be studied by the use of humanplasma clots and their lysis by the respective fibrinolytic plasminogenactivators.

Materials

Dog plasma is obtained from conscious mixed-breed hounds (either sexButler Farms, Clyde, N.Y., U.S.A.) by venipuncture into 3.8 percentcitrate. Fibrinogen is prepared from fresh dog plasma and humanfibrinogen is prepared from in-date ACD human blood at the fraction I-2according to previous procedures and specification. Smith, Biochem. J.,185, 1–11 (1980; and Smith, et al., Biochemistry, 11, 2958–2967, (1972).Human fibrinogen (98 percent pure/plasmin free) is from AmericanDiagnostica, Greenwich, Conn. Radiolabeling of fibrinogen I-2preparations is performed as previously reported. Smith, et al.,Biochemistry, 11, 2958–2967, (1972). Urokinase is purchased from LeoPharmaceuticals, Denmark, as 2200 Ploug units/vial. Streptokinase ispurchased from Hoechst-Roussel Pharmaceuticals, Somerville, N.J.

Methods—Effects on Lysis of Human Plasma Clots by t-PA

Human plasma clots are formed in micro test tubes by adding 50 μLthrombin (73 NIH unit/mL) to 100 μL human plasma which contains 0.0229μCi 125-iodine labeled fibrinogen. Clot lysis is studied by overlayingthe clots with 50 μL of urokinase or streptokinase (50, 100, or 1000unit/mL) and incubating for 20 hours at room temperature. Afterincubation the tubes are centrifuged in a Beckman Microfuge. 25 μL ofsupernate is added into 1.0 mL volume of 0.03 M tris/0.15 M NaCl bufferfor gamma counting. Counting controls 100 percent lysis are obtained byomitting thrombin (and substituting buffer). The factor Xa inhibitorsare evaluated for possible interference with fibrinolysis by includingthe compounds in the overlay solutions at 1, 5, and 10 μg/mLconcentrations. Rough approximations of IC₅₀ values are estimated bylinear extrapolations from data points to a value which would represent50 percent of lysis for that particular concentration of fibrinolyticagent.

Anticoagulant Activity

Materials

Dog plasma and rat plasma are obtained from conscious mixed-breed hounds(either sex, Butler Farms, Clyde, N.Y., U.S.A.) or from anesthetizedmale Sprague-Dawley rats (Harlan Sprague-Dawley, Inc., Indianapolis,Ind., U.S.A.) by venipuncture into 3.8 percent citrate. Fibrinogen isprepared from in-date ACD human blood as the fraction I-2 according toprevious procedures and specifications. Smith, Biochem. J., 185, 1–11(1980); and Smith, et al., Biochemistry, 11, 2958–2967 (1972). Humanfibrinogen is also purchased as 98 percent pure/plasmin free fromAmerican Diagnostica, Greenwich, Conn. Coagulation reagents Actin,Thromboplastin, Innovin and Human plasma are from Baxter HealthcareCorp., Dade Division, Miami, Fla. Bovine thrombin from Parke-Davis(Detroit, Mich.) is used for coagulation assays in plasma.

Methods

Anticoagulation Determinations

Coagulation assay procedures are as previously described. Smith, et al.,Thrombosis Research, 50, 163–174 (1988). A CoAScreener coagulationinstrument (American LABor, Inc.) is used for all coagulation assaymeasurements. The prothrombin time (PT) is measured by adding 0.05 mLsaline and 0.05 mL Thromboplastin-C reagent or recombinant human tissuefactor reagent (Innovin) to 0.05 mL test plasma. The activated partialthromboplastin time (APTT) is measured by incubation of 0.05 mL testplasma with 0.05 mL Actin reagent for 120 seconds followed by 0.05 mLCaCl₂ (0.02 M). The thrombin time (TT) is measured by adding 0.05 mLsaline and 0.05 mL thrombin (10 NIH units/mL) to 0.05 mL test plasma.The compounds of formula I are added to human or animal plasma over awide range of concentrations to determine prolongation effects on theAPTT, PT, and TT assays. Linear extrapolations are performed to estimatethe concentrations required to double the clotting time for each assay.Compounds of the instant invention potently extended the prolongationtimes in the APTT and PT assays, for example in some cases, with assayconcentrations necessary to double the APPT or PT of less than 1 μM.

Animals

Male Sprague Dawley rats (350–425 gm, Harlan Sprague Dawley Inc.,Indianapolis, Ind.) are anesthetized with xylazine (20 mg/kg, s.c.) andketamine (120 mg/kg, s.c.) or preferably are anesthetized usingisoflurane anesthesia (2–3%, conveniently 2.5%, for surgery; 1.5–2.5%,conveniently 2.5%, for maintenance; flow rate kept at 0.5% throughout)and maintained on a heated water blanket (37° C.). The jugular vein(s)is cannulated to allow for infusions.

Arterio-Venous Shunt Model

The left jugular vein and right carotid artery are cannulated with 20 cmlengths of polyethylene PE 60 tubing. A 6 cm center section of largertubing (PE 190) with a cotton thread (5 cm) in the lumen, is frictionfitted between the longer sections to complete the arterio-venous shuntcircuit. Blood is circulated through the shunt for 15 min before thethread is carefully removed and weighed. The weight of a wet thread issubtracted from the total weight of the thread and thrombus (see J. R.Smith, Br J Pharmacol, 77:29, 1982).

FeCl₃ Model of Arterial Injury

The carotid arteries are isolated via a midline ventral cervicalincision. A thermocouple is placed under each artery and vesseltemperature is recorded continuously on a strip chart recorder. A cuffof tubing (0.058 ID×0.077 OD×4 mm, Baxter Med. Grade Silicone), cutlongitudinally, is placed around each carotid directly above thethermocouple. FeCl₃ hexahydrate is dissolved in water and theconcentration (20 percent) is expressed in terms of the actual weight ofFeCl₃ only. To injure the artery and induce thrombosis, 2.85 μL ispipetted into the cuff to bathe the artery above the thermocouple probe.Arterial occlusion is indicated by a rapid drop in temperature. The timeto occlusion is reported in minutes and represents the elapsed timebetween application of FeCl₃ and the rapid drop in vessel temperature(see K. D. Kurz, Thromb. Res., 60:269, 1990).

Ex Vivo Coagulation Parameters

Ex vivo plasma thrombin time (TT), prothrombin time (PT) and activatedpartial thromboplastin time (APTT) are measured with a fibrometer. Bloodis sampled from a jugular catheter and collected in syringe containingsodium citrate (3.8 percent, 1 part to 9 parts blood). To measure TT,rat plasma (0.1 mL) is mixed with isotonic saline (0.1 mL) and bovinethrombin (0.1 mL, 30 U/mL in TRIS buffer; Parke Davis) at 37° C. For PT,to plasma (0.1 mL) mixed with isotonic saline (0.1 mL) is added PTreagent (0.1 mL, Dade, Thromboplastin-C); and the fibrometer startedimmediately after the addition of the final reagent. For APTT, plasma(0.1 mL) and APTT solution (0.1 mL, Organon Teknika) are incubated for 5minutes (37° C.); and CaCl₂ (0.1 mL, 0.025 M) is added to startcoagulation. Assays are done in duplicate and averaged.

Index of Bioavailability

Bioavailability studies may be conducted as follows. Compounds areadministered as aqueous solutions, or as solutions in 5% PEG 200, tomale Fisher rats, intravenously (iv) at 5 mg/kg via tail vein injectionand orally (po) as aqueous solutions, or as a suspension in 5% acacia,to fasted animals at 20 mg/kg by gavage. Serial blood samples areobtained at 5, 30, 120, and 240 minutes postdose following intravenousadministration and at 1, 2, 4, and 6 hours after oral dosing. Plasma isanalyzed for drug concentration using an HPLC procedure involving C8Bond Elute (Varian) cartridges for sample preparation and a methanol/30nM ammonium acetate buffer (pH 4) gradient optimized for each compound.% Oral bioavailability is calculated by the following equation:

${\%\mspace{14mu}{Oral}\mspace{14mu}{bioavailability}} = {\frac{{AUC}\mspace{14mu}{po}}{{AUC}\mspace{14mu}{iv}} \times \frac{{Dose}\mspace{14mu}{iv}}{{Dose}\mspace{14mu}{po}} \times 100}$where AUC is area under the curve calculated from the plasma level ofcompound over the time course of the experiment following oral (AUC po)and intravenous (AUC iv) dosing.Compounds

For oral determinations, the compound may be administered orally, bygavage, as a suspension in 5% acaia to conscious fasted rats. Thepretreatment time before flow is established through the shunt isselected based upon the peak apparent plasma concentration recorded inpreliminary time course experiments that track apparent drugconcentration in plasma following oral administration to consciousfasted rats, and typically varies between 1 to 5 hours. Animals used inantithrombotic efficacy experiments are anesthetized as described 15minutes befoe the predetermined pretreatment time to allow for surgicalpreparation of the animals. Compound solutions are prepared fresh dailyin normal saline or in 5% PEG200 in water for iv determinations and areinjected as a bolus or are infused starting 15 minutes before andcontinuing throughout the experimental perturbation which is 15 minutesin the arteriovenous shunt model and 60 minutes in the FeCl₃ model ofarterial injury and in the spontaneous thrombolysis model. Typically,bolus injection volume is 1 mL/kg for iv, and 5 mL/kg for po, andinfusion volume is 3 mL/h. For a similar procedure run in theanesthesized rabbit, for example an infusion rate of 6.8 mL/h was usedfor one compound infused in 5% PEG200 in water.

Statistics

Results are expressed as means+/− SEM. One-way analysis of variance isused to detect statistically significant differences and then Dunnett'stest is applied to determine which means are different. Significancelevel for rejection of the null hypothesis of equal means is P<0.05.

Animals

Male dogs (Beagles; 18 months-2 years; 12–13 kg, Marshall Farms, NorthRose, N.Y. 14516) are fasted overnight and fed Purina certifiedPrescription Diet (Purina Mills, St. Louis, Mo.) 240 minutes afterdosing. Water is available ad libitum. The room temperature ismaintained between 66–74° F.; 45–50 percent relative humidity; andlighted from 0600–1800 hours.

Pharmacokinetic Model.

Test compound is formulated immediately prior to dosing by making asuspension in a “wet granulaion” (povidone, 0.85 mg/mL; lactose, 15.0mg/mL; and polysorbate 80, 65 μL in 250 mL water). Dogs are given asingle 20 mg/kg (in 25 mL of wet granulation) dose of test compound byoral gavage. Blood samples (4.5 mL) are taken from the cephalic vein at0.25, 0.5, 0.75, 1, 2, 3, 4 and 6 hours after dosing. Samples arecollected in citrated Vacutainer tubes and kept on ice prior toreduction to plasma by centrifugation. Plasma samples are analyzed byHPLC MS. Plasma concentration of test compound is recorded and used tocalculate the pharmacokinetic parameters: elimination rate constant, Ke;total clearance, Clt; volume of distribution, VD; time of maximum plasmatest compound concentration, Tmax; maximum concentration of testcompound of Tmax, Cmax; plasma half-life, t0.5; and area under thecurve, A.U.C.; fraction of test compound absorbed, F.

Canine Model of Coronary Artery Thrombosis

Male dogs (Beagles, as described above) are fasted overnight and dosedwith test compound that is fomulated immediately prior to dosing bymaking a suspension in a “wet granulation” as described above. Dogs aregiven a single dose of 5, 10 or 20 mg/kg (in 25 mL of wet granulation)of test compound by oral gavage. Based on the pharmacokinetics of thetest compound, dogs are dosed either 1 or 2 hours prior to anesthesia.Dogs are anesthetized with sodium pentobarbital (30 mg/kg intravenouslyi.v.), intubated, and ventilated with room air. Tidal volume andrespiratory rates are adjusted to maintain blood PO₂, PCO₂, and pHwithin normal limits. Subdermal needle electrodes are inserted for therecording of a lead II ECG.

The left jugular vein and common carotid artery are isolated through aleft mediolateral neck incision. Arterial blood pressure (ABP) ismeasured continuously with a precalibrated Millar transducer (modelMPC-500, Millar Instruments, Houston, Tex., U.S.A.) inserted into thecarotid artery. The jugular vein is cannulated for blood sampling duringthe experiment. In addition, the femoral veins of both hindlegs arecannulated for administration of test compound.

A left thoracotomy is performed at the fifth intercostal space, and theheart is suspended in a pericardial cradle. A 1- to 2-cm segment of theleft circumflex coronary artery (LCX) is isolated proximal to the firstmajor diagonal ventricular branch. A 26-gauge needle-tipped wire anodalelectrode (Teflon-coated, 30-gauge silverplated copper wire) 3–4 mm longis inserted into the LCX and placed in contact with the intimal surfaceof the artery (confirmed at the end of the experiment). The stimulatingcircuit is completed by placing the cathode in a subcutaneous (s.c.)site. An adjustable plastic occluder is placed around the LCX, over theregion of the electrode. A precalibrated electromagnetic flow probe(Carolina Medical Electronics, King, N.C., U.S.A.) is placed around theLCX proximal to the anode for measurement of coronary blood flow (CBF).The occluder is adjusted to produce a 40–50 percent inhibition of thehyperemic blood flow response observed after 10-s mechanical occlusionof the LCX. All hemodynamic and ECG measurements are recorded andanalyzed with a data acquisition system (Notochord HEM data analysissystem, Croissy, France).

Thrombus Formation and Compound Administration Regimens

Electrolytic injury of the intima of the LCX is produced by applying100-μA direct current (DC) to the anode. The current is maintained for60 min and then discontinued whether the vessel has occluded or not.Thrombus formation proceeds spontaneously until the LCX is totallyoccluded (determined as zero CBF and an increase in the S-T segment fora minimum of 30 minutes). The preparation is followed for 4 hours atwhich time the animal is euthanized and the thrombus is dissected fromthe LCX and weighed.

Hematology, Coagulation and Template Bleeding Time Determinations

Citrated blood (3 mL, 1 part 3.8% citrate: 9 parts blood) is drawnbefore drug administration, at 60 min after administration, at 60 minafter initiation of vessel injury and just prior to the end of theexperiment. Whole blood cell counts, hemoglobin, and hematocrit valuesare determined on a 40-μL sample of the citrated whole blood with ahematology analyzer (Cell-Dyn 900, Sequoia-Turner, Mount View, Calif.,U.S.A.). The remaining blood was cetrifuged at 3,000 g for 5 min toprepare cell-free plasma. Plasma clotting times, prothrombin time (PT)and activated partial thromoplastin times (APTT) were performed usingstandard Dade reagents and the Coa-Screener coagulation device (AmericanLabor, Largo, Fla.). Gingival template bleeding times are determinedwith a Simplate II bleeding time device (organon Teknika Durham, N.C.,U.S.A.). The device is used to make 2 horizontal incisions in thegingiva of either the upper or lower left jaw of the dog. Each incisionis 3 mm wide×2 mm deep. The incisions are made, and a stopwatch is usedto determine how long bleeding occurs. A cotton swab is used to soak upthe blood as it oozes from the incision. Template bleeding time is thetime from incision to stoppage of bleeding. Bleeding times are takenjust before administration of test compound (0 min), 60 min intoinfusion, at conclusion of administration of the test compound (120min), and at the end of the experiment.

All data are analyzed by one-way analysis of variance (ANOVA) followedby Dunnet's post hoc t test to determine the level of significance.Repeated-measures ANOVA are used to determine significant differencesbetween time points during the experiments. Values are determined to bestatistically different at least at the level of p<0.05. All values aremean±SEM. All studies are conducted in accordance with the guidingprinciples of the American Physiological Society. Further detailsregarding the procedures are described in Jackson, et al., J.Cardiovasc. Pharmacol., (1993), 21, 587–599.

Compounds of the instant invention are potent anticoagulant andantithrombotic agents which exhibit particularly good plasma exposurefollowing oral administration, as well as desirable volume ofdistribution and tissue selectivity properties, as evidenced by standardpharmacokinetic/pharmcodynamic and brain flux assays.

The following Examples are provided to further describe the inventionand are not to be construed as limitations thereof.

The abbreviations, symbols and terms used in the examples have thefollowing meanings.

-   -   Ac=acetyl    -   Analysis=elemental analysis    -   aq=aqueous    -   Boc=t-butyloxycarbonyl    -   Calcd=calculated    -   conc=concentrated    -   DMF=dimethylformamide    -   DMSO=dimethylsulfoxide    -   EtOAc=ethyl acetate    -   EtOH=ethanol    -   MeOH=methanol    -   HPLC=High Performance Liquid Chromatography    -   IR=Infrared Spectrum    -   APCI-MS=atmospheric pressure chemical ionization mass spectrum    -   ESI-MS (or ES-MS)=electrospray ionization mass spectrum    -   FD-MS=field desorption mass spectrum    -   IS-MS=ion spray mass spectrum    -   NMR=Nuclear Magnetic Resonance    -   RPHPLC=Reversed Phase High Performance Liquid Chromatography    -   RT (or Rt)=retention time    -   satd=saturated    -   SCX=strong cation exchange (resin)    -   TFA=trifluoroacetic acid    -   THF=tetrahydrofuran

Unless otherwise stated, pH adjustments and work up are with aqueousacid or base solutions. ¹H-NMR indicates a satisfactory NMR spectrum wasobtained for the compound described. IR indicates a satisfactory infrared spectrum was obtained for the compound described.

Analytical HPLC method was a linear gradient of 90/10 to 50/50 (0.1% TFAin water/0.1% TFA in acetonitrile) over 40 minutes with a flow rate of 1mL/min.

PREPARATION OF NITRO-AMIDE (NA) INTERMEDIATES

General Procedure NA-A:

2-Nitro-N-(5-methylpyridin-2-yl)benzamide.

To a stirring solution of 2-amino-5-methylpyridine (3.1 g, 29 mmol) indichloromethane (200 mL) was added pyridine (7.3 mL, 90 mmol) followedby 2-nitrobenzoyl chloride (5.7 g, 30 mmol). After 4 h, the solvent wasremoved in vacuo and the residue was partitioned between ethyl acetate(500 mL) and water (250 mL). The organic phase was separated and washedwith water, brine, dried (MgSO₄) and filtered, and then concentrated invacuo to a volume of about 100 mL (precipitate observed). The mixturewas then sonicated and allowed to stand overnight then filtered. Thecollected solid was then washed with diethyl ether, filtered and driedunder vacuum to give 3.9 g (52%) of the title compound.

¹H-NMR

FD-MS, m/e 256.9 (M+).

Analysis for C₁₃H₁₁N₃O₃: Calc: C, 60.70; H, 4.31; N, 16.33; Found: C,61.21; H, 4.32; N, 16.63.

General Procedure NA-B:

4-Chloro-N-(5-chloropyridin-2-yl)-2-nitrobenzamide.

To a stirring suspension of 4-chloro-2-nitrobenzoic acid (20 g, 99 mmol)in dichloromethane (500 mL) was added a few drops of DMF, followed byoxalyl chloride (15.1 g, 119 mmol). After 1.h, the solvent was removedin vacuo and the residue was dissolved in dichloromethane (500 mL). Tothis stirring solution was added pyridine (24 mL, 297 mmol) followed by2-amino-5-chloropyridine (12.7 g, 99 mmol). After stirring overnight,the solvents were removed in vacuo and the residue was stirredvigorously with ethyl acetate and water for several hours. The mixturewas filtered to give a white solid, which was washed with ethyl acetateand dried in vacuo to give 23 g (74%) of the title compound. Thecombined ethyl acetate washings and extract were then washed twice with1 M citric acid, once with brine, twice with saturated aq sodiumbicarbonate, and again with brine. The organic phase was then dried withMgSO₄, filtered and concentrated in vacuo. The solid was then suspendedin diethyl ether, sonicated and filtered to give a second crop of thetitle compound as a white solid (5.79 g, 19%).

¹H-NMR

IS-MS, m/e 312.0 (M+1)

Analysis for C₁₂H₇N₃O₃Cl₂: Calcd: C, 46.18;.H, 2.26; N, 13.46; Found: C,46.24; H, 2.37; N, 13.43.

Preparation of Intermediates NA-1-NA-12

The following exemplary nitro-amide intermediates were prepared using aconventional procedure such as that of General Procedure NA-A or GeneralProcedure NA-B, or as otherwise described.

Intermediate NA-1

5-Fluoro-2-nitro-N-(5-fluoropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 278.09 (M−1).

The starting 4-fluoro-2-nitrobenzoic acid was prepared as follows:

To a stirring solution of KMnO₄ (76 g, 483 mmol) in water (1 L) wasadded 4-fluoro-2-nitrotoluene and the solution was heated to reflux.After 4 h, the hot mixture was filtered and the filtrate was cooled withice, washed with diethyl ether, acidified with conc HCl, and thenextracted twice with diethyl ether. The combined ether extracts werewashed with brine, dried with MgSO₄, filtered and concentrated in vacuoto give 12.07 g (34%) of a white solid.

¹H-NMR

IS-MS, m/e 184.0 (M−1).

Analysis for C₇H₄NO₄F: Calcd: C, 45.42; H, 2.18; N, 7.57; Found: C,45.63; H, 2.30; N, 7.61.

Intermediate NA-2

5-Fluoro-2-nitro-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 296.22 (M+1).

Analysis for C₁₂H₇ClFN₃O₃: Calcd: C,48.75; H,2.39; N,14.21; Found:C,48.57; H,2.37; N,14.19.

Intermediate NA-3

5-Chloro-2-nitro-N-(5-fluoropyridin-2-yl)benzamid.

¹H-NMR

ESI-MS, m/e 296.24 (M+1).

Analysis for C₁₂H₇ClFN₃O₃: Calcd: C, 48.75; H, 2.39; N, 14.21; Found: C,48.97; H, 2.61; N, 14.13.

Intermediate NA-4

5-Chloro-2-nitro-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 311.96 (M+1).

Analysis for C₁₂H₇Cl₂N₃O₃: Calcd: C, 46.18; H, 2.26; N, 13.46; Found: C,46.24; H, 2.22; N, 13.29.

Intermediate NA-5

5-Chloro-2-nitro-N-(5-methylpyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 291.97 (M+1).

Analysis for C₁₃H₁₀ClN₃O₃: Calcd: C, 53.53; H, 3.46; N, 14.41; Found: C,53.76; H, 3.41; N, 14.35.

Intermediate NA-6

5-Methyl-2-nitro-N-(5-fluoropyridin-2-yl)benzamide.

¹H-NMR

APCI-MS, m/e 276 (M+1).

Intermediate NA-7

5-Methyl-2-nitro-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 292 (M+1).

Analysis for C₁₃H₁₀ClN₃O₃: Calcd: C, 53.53; H, 3.46; N, 14.41; Found: C,53.52; H, 3.56; N, 14.49.

Intermediate NA-8

5-Methyl-2-nitro-N-(5-methylpyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 272.37 (M+1).

Intermediate NA-9

4,5-Difluoro-2-nitro-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 313.95 (M+1).

Analysis for C₁₂H₆ClF₂N₃O₃: Calcd: C, 45.95; H, 1.93; N, 13.40; Found:C, 45.77; H, 2.00; N, 13.43.

Intermediate NA-10 Compound AA-10 prepared via4-amino-3-iodoacetophenone Intermediate NA-11

4-Methoxycarbonyl-2-nitro-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 336.09 (M+1).

Analysis for C₁₄H₁₀ClN₃O₅: Calcd: C, 50.09; H, 3.00; N, 12.52; Found: C,49.83; H, 3.08; N, 12.25.

Intermediate NA-12

5-Methoxycarbonyl-2-nitro-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 336.07 (M+1).

Analysis for C₁₄H₁₀ClN₃O₅: Calcd: C, 50.09; H, 3.00; N, 12.52; Found: C,50.37; H, 3.08; N, 12.52.

Preparation of Amino-Amide (AA) Intermediates

General Procedure AA-A:

N-(5-Methylpyridin-2-yl)-2-aminobenzamide.

To a stirring solution of N-(5-methylpyridin-2-yl)-2-nitrobenzamide (1.5g, 5.8 mmol) and Ni(OAc)₂.4H₂O (2.9 g, 11.7 mmol) in THF (20 mL) andmethanol (40 mL) at 0° C. was added, in small portions, sodiumborohydride (0.88 g, 23.2 mmol). After complete addition and anadditional 5 min, the solvent was evaporated in vacuo and the residuewas partitioned between ethyl acetate (200 mL) and 50% conc NH₄OH (206mL). The organic phase was separated and washed again with 50% concNH₄OH, followed by brine, then dried with MgSO₄, filtered andconcentrated in vacuo to give 1.25 g (95%) of a light yellow solid.

¹H-NMR

FD-MS, m/e 227.1 (M+).

General Procedure AA-B:

N-(5-Chloropyridin-2-yl)-2-aminobenzamide.

To a solution of N-(5-chloropyridin-2-yl)-2-nitrobenzamide (2 g, 7.2mmol) in THF (50 mL) and ethyl acetate (50 mL) was added Raney Ni (0.2g) and the mixture was placed under hydrogen (4.1 bar) in a highpressure apparatus. After shaking overnight, the mixture was filteredand concentrated in vacuo and purified by flash chromatography to give1.5 g (83%) of an off-white solid.

¹H-NMR

Preparation of Intermediates AA-1-AA-14

The following exemplary amino-amide intermediates were prepared using aconventional procedure such as that of General Procedure AA-A or GeneralProcedure AA-B, or as otherwise described.

Intermediate AA-1

2-Amino-5-fluoro-N-(5-fluoropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 248 (M−1)

Intermediate AA-2

2-Amino-5-fluoro-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 264.17 (M−1).

Analysis for C₁₂H₉ClFN₃O: Calcd: C, 54.25; H, 3.41; N, 15.82; Found: C,53.96; H, 3.43; N, 15.54.

Intermediate AA-3

2-Amino-5-chloro-N-(5-fluoropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 264.13 (M−1).

Intermediate AA-4

2-Amino-5-chloro-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 281.12 (M−1).

Analysis for C₁₂H₉Cl₂N₃O: Calcd: C, 51.09; H, 3.22; N, 14.89; Found: C,51.19; H, 3.33; N, 14.61.

Intermediate AA-5

2-Amino-5-chloro-N-(5-methylpyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 260.02 (M−1).

Analysis for C₁₃H₁₂ClN₃O: Calcd: C, 59.66; H, 4.62; N, 16.06; Found: C,59.89; H, 4.57; N, 15.99.

Intermediate AA-6

2-Amino-5-methyl-N-(5-fluoropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 244.41 (M−1).

Intermediate AA-7

2-Amino-5-methyl-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 260.01 (M−1).

Intermediate AA-8

2-Amino-5-methyl-N-(5-methylpyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 240.15 (M−1).

Intermediate AA-9

2-Amino-4,5-difluoro-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 282.12 (M−1).

Intermediate AA-10

5-Acetyl-2-amino-N-(5-chloropyridin-2-yl)benzamide.

To a stirred solution of 4-aminoacetophenone (50 g, 370 mmol) in ethanol(1 L) and dichloromethane (750 mL) at 0° C. was added iodine (94 g, 370mmol) and silver sulfate (116 g, 370 mmol). The reaction mixture wasstirred at 0° C. for 10 min and then at room temperature for 4 h,filtered and concentrated. The resulting residue was partitioned betweendichloromethane and 5 N sodium hydroxide. The organic phase was dried(magnesium sulfate), filtered, and concentrated in vacuo. The crudeproduct was chromatographed over silica gel, eluting with a stepwisegradient from dichloromethane to 10% ethyl acetate in dichloromethane,to give 38.2 g (40%) of 4-amino-3-iodoacetophenone as a yellow oil.

A mixture of 4-amino-3-iodoacetophenone (5.0 g, 19.2 mmol),2-amino-5-chloropyridine (7.4 g, 54.5 mmol), palladium acetate (431 mg,1.92 mmol), 1,3-bis(diphenylphosphino)propane (2.37 g, 5.75 mmol),triethylamine (5.4 mL, 38.3 mmol) in acetonitrile (100 mL) was shakenunder a carbon monoxide atmosphere (54.4 bar) for 16 h. The crudemixture was filtered through diatomaceous earth, and the filtrate wasstripped to near dryness and titurated with ethyl ether to remove theexcess 2-amino-5-chloropyridine. The product was isolated by SCX column,then flash chromatography using 10% acetonitrile in chloroform. Thisafforded 1.17 g (21%) of the title product.

¹H-NMR

ESI-MS, m/e 288.1 (M−1).

Intermediate AA-11

2-Amino-4-methoxycarbonyl-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 304.09 (M−1).

Intermediate AA-12

2-Amino-5-methoxycarbonyl-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 304 (M−1).

Intermediate AA-13

3-Amino-N-(5-chloropyridin-2-yl)pyridine-2-carboxamide.

¹H-NMR

ESI-MS, m/e 247.19 (M−1).

The amine was prepared using a similar procedure to the following:

A (Parr) pressure apparatus was charged with 3-amino-2-chloropyridine(500 mg, 3.89 mmol), 2-amino-5-chloropyridine (1.00 g, 7.78 mmol),palladium acetate (88 mg, 0.39 mmol), 1,3-bis(diphenylphosphino)propane(483 mg, 1.17 mmol) and triethylamine (590 mg, 5.84 mmol). The mixturewas placed under a carbon monoxide atmosphere (4.1 bar) and heated at100° C. After 72 h, the mixture was filtered, concentrated and theresidue purified by column chromatography (SiO₂: 0 to 5% EtOAc inmethylene chloride) affording 550 mg (57%) of the title compound.

¹H-NMR, IR

IS-MS, m/e 249 (M+1).

Analysis for C₁₁H₉ClN₄O: Calcd: C, 53.13; H, 3.65; N, 22.53; Found: C,53.40; H, 3.66; N, 22.45.

Intermediate AA-14

3-Amino-N-(5-chloropyridin-2-yl)-6-methylpyridine-2-carboxamide.

Using methods substantially equivalent to those described above forIntermediate AA-13,3-amino-N-(5-chloropyridin-2-yl)-6-methylpyridine-2-carboxamide (16 g,46%) was prepared from 3-amino-2-chloro-6-methylpyridine and2-amino-5-chloropyridine.

¹H NMR

ESI-MS, m/e 263.1 (M+1).

Preparation of Intermediates A-1–A-5

The following intermediates were prepared by acylation of the requisiteamine using 4-fluorobenzoyl chloride and a procedure similar to thatdescribed for the preparation of Intermediate A-2, or as otherwisedescribed.

Intermediate A-1

5-Fluoro-2-(4-fluorobenzoylamino)-N-(5-chloropyridin-2-yl)-benzamide.

¹H-NMR (DMSO) δ 8.43 (d, J=2.6 Hz), 8.13 (d, J=9.0 Hz), 8.11 (d, J=9.0Hz), 7.93–7.99 (m), 7.71 (dd, J=9.4 Hz and 3.0 Hz), 7.35–7.50 (m).

ESI-MS, m/e 388.09 (M+1).

Analysis for C₁₉H₁₂ClF₂N₃O₂: Calcd: C, 58.85; H, 3.12; N, 10.84; Found:C, 58.64; H, 3.16; N, 10.82.

Intermediate A-2

5-Chloro-2-(4-fluorobenzoylamino)-N-(5-chloropyridin-2-yl)-benzamide(General Procedure).

To a solution of 2-amino-5-chloro-N-(5-chloropyridin-2-yl)benzamide(2.82 g, 10 mmol) in methylene chloride (100 mL) and pyridine (3 mL) wasadded 4-fluorobenzoyl chloride (1.42 mL, 12 mmol) dropwise at 0° C. Thereaction mixture was warmed to room temperature and stirred overnight.Water (50 mL) was added; and the mixture was stirred for 30 min beforethe solids were filtered, washed with saturated sodium bicarbonate (30mL), water (30 mL), and ether (50 mL), and dried under vacuum to providethe product (3.2 g, 79%) pure enough to use in the next step.

¹H-NMR (DMSO, 250 MHz) δ 8.46 (d, J=2.1 Hz), 8.11–8.20 (m), 7.93–8.01(m), 7.68 (dd, J=8.8 Hz and 2.4 Hz), 7.37–7.44 (m).

ESI-MS, m/e 402.17 (M−1).

Analysis for C₁₉H₁₂Cl₂FN₃O₂: Calcd: C, 56.46; H, 2.99; N, 10.40; Found:C, 55.97; H, 2.93; N, 10.43.

Int rmediate A-3

5-Chloro-2-(4-fluorobenzoylamino)-N-(5-methylpyridin-2-yl)-benzamide.

¹H-NMR (DMSO) δ 8.31 (d, J=9.14 Hz), 8.23 (d, J=1.8), 7.95–7.99 (m),7.64–7.69 (m), 7.37–7.43 (m), 2.28 (s).

ESI-MS, m/e 384.21 (M+1).

Analysis for C₂₀H₁₅ClFN₃O₂: Calcd: C, 62.59; H, 3.94; N, 10.95; Found:C, 62.36; H, 4.04; N, 10.80.

Intermediate A-4

2-(4-Fluorobenzoylamino)-5-methyl-N-(5-methylpyridin-2-yl)-benzamide.

¹H-NMR (DMSO) δ 8.1 (d, J=8.1 Hz), 7.88–7.93 (m), 7.75 (s), 7.60 (d,J=8.4 Hz), 7.30–7.36 (m), 2.29 (s), 2.22 (s).

FD-MS, m/e 364.21 (M+1).

Analysis for C₂₁H₁₈FN₃O₂: Calcd: C, 69.41; H, 4.99; N, 11.56; Found: C,69.03; H, 4.85; N, 11.51.

Intermediate A-5

3-(4-Fluorobenzoylamino)-N-(5-chloropyridin-2-yl)pyridine-2-carboxamide.

¹H-NMR (DMSO) δ 8.14 (dd, J=8.4 Hz and 1.5 Hz), 8.46 (d, J=2.9 Hz), 8.23(d, J=8.8 Hz), 8.02–8.06 (m), 7.75–7.79 (2d, J₁=4.4 Hz, J₂=4.8 Hz),7.44–7.50 (m), 7.29–7.35 (m)

ESI-MS, m/e 369.17 (M−1).

Analysis for C₁₈H₁₂ClFN₄O₂: Calcd: C, 58.31; H, 3.26; N, 15.11; Found:C, 57.82; H, 2.99; N, 14.95.

Preparation of Intermediates B-1–B-2

The following intermediates were prepared by acylation of the requisiteamine using 4-fluoro-2-trifluoromethyl-benzoyl chloride and a proceduresimilar to that described for the preparation of Intermediate A-2, or asotherwise described.

Intermediate B-1

5-Fluoro-2-(4-fluoro-2-trifluoromethylbenzoylamino)-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR (DMSO) δ 8.44 (d, J=2.3 Hz), 8.14 (d, J=8.7 Hz), 7.93 (dd, J=9.0Hz and 2.6 Hz), 7.66–7.86 (m), 7.59 (dd, J=9.1 Hz and 2.8 Hz), 7.41–7.48(m).

ESI-MS; m/e 456.20 (M+1).

Intermediate B-2

5-Chloro-2-(4-fluoro-2-trifluoromethylbenzoylamino)-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR (DMSO) δ 8.44 (d, J=2.3 Hz), 8.12 (d, J=9.0 Hz), 7.92 (dd, J=9.0Hz and 2.6 Hz), 7.63–7.87 (m).

ESI-MS, m/e 472.14 (M+1).

Analysis for C₂₀H₁₁Cl₂F₄N₃O₂: Calcd: C, 50.87; H, 2.35; N, 8.90; Found:C, 50.58; H, 2.30; N, 8.61.

Preparation of Intermediates C-1–C-2

The following intermediates were prepared by acylation of the requisiteamine using 4-fluoro-3-trifluoromethylbenzoyl chloride and a proceduresimilar to that described for the preparation of Intermediate A-2, or asotherwise described.

Intermediate C-1

5-Fluoro-2-(4-fluoro-3-trifluoromethylbenzoylamino)-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR (DMSO) δ 8.39 (d, J=2.6 Hz), 8.25 (dd, J=6.9 Hz and 2.6 Hz), 8.12(d, J=8.7 Hz), 7.87–7.95 (m), 7.62–7.73 (m), 7.74–7.50 (m).

ESI-MS, m/e 456.19 (M+1).

Analysis for C₂₀H₁₁ClF₅N₃O₂: Calcd: C, 52.71; H, 2.43; N, 9.22; Found:C, 52.97; H, 2.33; N, 9.01.

Intermediate C-2

5-Chloro-2-(4-fluoro-3-trifluoromethylbenzoylamino)-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR (DMSO) δ 8.41 (d, J=2.6 Hz), 8.24 (d, J=6.8 Hz), 8.11 (d, J=9.0Hz), 7.92–7.98 (m), 7.86 (d, J=2.3 Hz), 7.64–7.74 (m)

ESI-MS, m/e 472.11 (M+1).

Analysis for C₂₀H₁₁Cl₂F₄N₃O₂: Calcd: C, 50.87; H, 2.35; N, 8.90; Found:C, 51.02; H, 2.16; N, 8.69.

Preparation of Intermediate D-1

The following intermediate was prepared by acylation of the requisiteamine using 3,4-difluorobenzoyl chloride and a procedure similar to thatdescribed for the preparation of Intermediate A-2, or as otherwisedescribed.

Intermediate D-1

2-(3,14-Difluorobenzoylamino)-5-fluoro-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR (DMSO) δ 8.42 (d, J=2.6 Hz), 8.11 (d, J=9.0 Hz), 7.88–8.02 (m),7.77 (s, br), 7.58–7.70 (m), 7.43–7.50 (m).

ESI-MS, m/e 406.35 (M+1).

Preparation of Intermediates E-1–E-2

The following intermediates were prepared by acylation of the requisiteamine using 3,4,5-trifluorobenzoyl chloride and a procedure similar tothat described for the preparation of Intermediate A-2, or as otherwisedescribed.

Intermediate E-1

5-Fluoro-2-(3,4,5-trifluorobenzoylamino)-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR (DMSO) δ 8.41 (d, J=2.6 Hz), 8.12 (d, J=9.0 Hz), 7.77–7.97 (m),7.65 (dd, J=9.3 Hz and 2.8 Hz), 7.43–7.50 (m).

ESI-MS, m/e 423.99 (M+1).

Intermediate E-2

5-Chloro-2-(3,4,5-trifluorobenzoylamino)-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR (DMSO) δ 8.42 (d, J=2.3 Hz), 8.11 (d, J=9.0 Hz), 7.95 (dd, J=9.0Hz and 2.2 Hz), 7.79–7.87 (m), 7.66 (dd, J=9.0 Hz and 2.3 Hz).

ESI-MS, m/e 440.01 (M+1).

Analysis for C₁₉H₁₀Cl₂F₃N₃O₂: Calcd: C, 51.84; H, 2.29; N, 9.55; Found:C, 52.08; H, 2.28; N, 9.41.

Preparation of Intermediate F-1

5-Fluoro-2-(4-fluoro-2-hydroxybenzoylamino)-N-(5-chloropyridin-2-yl)benzamide.

A.5-Fluoro-2-(4-fluoro-2-acetoxybenzoylamino)-N-(5-chloropyridin-2-yl)benzamide.

2-Amino-5-fluoro-N-(5-chloropyridin-2-yl)benzamide (5.32 g, 20 mmol) wasadded to a solution of 2 mL dry pyridine and 2-acetoxy-4-fluorobenzoylchloride (21 mmol) in 100 mL dry methylene chloride under dry nitrogenwith magnetic stirring. The reaction mixture was allowed to stirovernight at room temperature and then was diluted with 500 mL ofmethylene chloride. The methylene chloride solution was washed with cold1 M HCl, cold saturated NaHCO₃, and brine; and then was dried oversodium sulfate. The solvent was evaporated, and the crude product wascrystallized from acetone to give 2.92 g of off-white product.

¹H-NMR

ESI-MS, m/e 446.1 (M+1).

B.5-Fluoro-2-(4-fluoro-2-hydroxybenzoylamino)-N-(5-chloropyridin-2-yl)benzamide.

The acetoxy compound from Part A above (2.75 g, 6 mmol) was dissolved in75 mL of methanol under nitrogen. The solution was cooled in anice-water bath and 0.5 M NaOH (13 mL, 6.5 mmol) was added. The reactionwas stirred overnight while warming to room temperature. One molar HCl(6.5 mL, 6.5 mmol) was added. The mixture was diluted with additionalwater and the precipitated solid was filtered, washed with water, anddried over KOH in vacuo at 35° C. to give 2.15 g of the title product asa white solid (89% yield).

¹H-NMR

ESI-MS, m/e 404.4 (M+1).

Analysis for C₂₁H₁₄ClF₂N₃O₄: Calcd: C, 56.52; H, 3.00; N, 10.41; Found:C, 56.34; H, 2.93; N, 9.99.

Preparation of Intermediates F-2–F-6

The following intermediates were prepared by alkylation of5-fluoro-2-(4-fluoro-2-hydroxybenzoylamino)-N-(5-chloropyridin-2-yl)benzamide(Intermediate F-1) using the described alkylating agent and a proceduresimilar to that described for the preparation of Intermediate F-2, or asotherwise described.

Intermediate F-2

5-Fluoro-2-(4-fluoro-2-ethoxybenzoylamino)-N-(5-chloropyridin-2-yl)-benzamide.

In a 4 mL screw cap vial fitted with a small egg-shaped stir bar, werecombined5-fluoro-2-(4-fluoro-2-hydroxybenzoylamino)-N-(5-chloropyridin-2-yl)benzamide(200 mg, 0.49 mmol) and powdered anhydrous K₂CO₃ (103 mg, 0.74 mmol, 1.5eq) in 0.5 mL of dry dimethylformamide (DMF). Ethyl iodide (115 mg, 0.74mmol) was then added, the vial was capped, and the slurry heated to 50°C. with vigorous stirring overnight. The slurry was diluted with 20 mLwater and extracted into 15 mL of dichloromethane. The organic layer wasthen washed with 20 mL portions of saturated NaHCO₃, water, and brine;then dried over Na₂SO₄ and concentrated in vacuo. The residue wasdissolved in a few mL of dichloromethane and purified by radialchromatography, eluting using hexanes, then 4:1 hexanes:EtOAc, toprovide 135 mg (63%) of the title compound as pinkish-white crystals.

ES-MS, m/e 432.1 (M+1).

Intermediate F-3

5-Fluoro-2-(4-fluoro-2-isopropoxybenzoylamino)-N-(5-chloropyridin-2-yl)benzamide.

Prepared using 2-iodopropane.

ES-MS, m/e 446.1 (M+1).

Intermediate F-4

5-Fluoro-2-[4-fluoro-2-(2-hydroxyethoxy)benzoylamino]-N-(5-chloropyridin-2-yl)benzamide.

Prepared using 2-bromoethanol and the addition of a few mg oftetrabutylammonium iodide and heating for 48 hours at 75° C. instead of36–40 hours at 50° C.

ES-MS, m/e 448.0 (M+1).

Intermediate F-5

5-Fluoro-2-[4-fluoro-2-(2-methoxyethoxy)benzoylamino]-N-(5-chloropyridin-2-yl)benzamide.

Prepared (46% yield) using 2-bromoethyl methyl ether and thetetrabutylammonium iodide and conditions as described for IntermediateF-4.

ES-MS, m/e 462.1 (M+1).

Intermediate F-6

5-Fluoro-2-[4-fluoro-2-(2-methylthioethoxy)benzoylamino]-N-(5-chloropyridin-2-yl)benzamide.

In a flame-dried 50 mL round bottomed single necked flask were combined5-fluoro-2-(4-fluoro-2-hydroxybenzoylamino)-N-(5-chloropyridin-2-yl)benzamide(491 mg, 1.22 mmol), 2-(methylthio)ethanol (160 μL, 168 mg, 1.82 mmol,1.5 eq), and triphenylphosphine (477 mg, 1.82 mmol) under a nitrogenatmosphere in 30 mL of dry tetrahydrofuran (THF) to produce amilky-white slurry. Diisopropyl azodicarboxylate (DIAD, 358 μL, 368 mg,1.82 mmol) was then added dropwise via syringe over 10–15 min to thestirring slurry. The milky-white slurry changed to a clear lightorange-yellow solution during DIAD addition and this solution wasstirred overnight under nitrogen at room temperature. The reactionmixture was concentrated in vacuo and the resulting orange oil wasdissolved in ethyl acetate (100 mL), washed with water and brine, thendried over sodium sulfate and concentrated again to yield an orange oil.The oil was purified on a 75 mL column of silica gel using a stepgradient of 100:0, 95:5, 9:1, 85:15 and 8:2 hexanes:EtOAc, then byradial chromatography with 9:1 and 8:2 hexanes:EtOAc and finally byrecrystallization from 9:1 hexanes:EtOAc to produce 213 mg (37%) of thetitle compound as white needles.

¹H-NMR

ESI-MS, m/e 478.1 (M+1)

PREPARATION OF EXAMPLES P1–P3

The following protected examples were prepared using1-t-butoxycarbonylhexahydro-1,4-diazepine and the requisite intermediateand a procedure similar to that described for the preparation of Example5, or as otherwise described.

EXAMPLE P1

2-[4-(4-t-Butoxycarbonylhexahydro-1,4-diazepin-1-yl)benzoylamino]-5-fluoro-N-(5-chloropyridin-2-yl)benzamide.

Directly Deprotected without Obtaining Spectra. EXAMPLE P2

2-[4-(4-t-Butoxycarbonylhexahydro-1,4-diazepin-1-yl)benzoylamino]-5-chloro-N-(5-chloropyridin-2-yl)benzamide.

Directly Deprotected without Obtaining Opectra.) EXAMPLE P3

2-[4-(4-t-Butoxycarbonylhexahydro-1,4-diazepin-1-yl)benzoylamino]-5-methyl-N-(5-methylpyridin-2-yl)benzamide.

Directly Deprotected without Obtaining Spectra. PREPARATION OF EXAMPLES1–3

The following examples were prepared using the requisite1-t-butoxycarbonyl protected hexahydro-1,4-diazepine described above anda procedure similar to that described for the preparation of Example 1,or as otherwise described.

Preparation of Salts

TFA salts were obtained by removal of the solvent after the deprotectionof nitrogen in methylene chloride and TFA.

Monohydrochlorides were prepared by treating the free bases suspended inmethylene chloride, methanol mixture (4:1) with one equivalent of HCl inether and sonicating for five minutes and evaporating off the solvent.

Similarly the polyhydrochlorides were prepared by treatment with excessHCl in ether.

EXAMPLE 1

5-Fluoro-2-[4-(hexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-chloropyridin-2-yl)benzamide.

To a suspension of2-[4-(4-t-butoxycarbonylhexahydro-1,4-diazepin-1-yl)benzoylamino]-5-fluoro-N-(5-chloropyridin-2-yl)benzamide(Example P1, 0.59 g) in methylene chloride (3 mL) and anisole (1 mL) wasadded trifluoroacetic acid (3 mL) dropwise. The mixture was stirredovernight at room temperature before the solvent was evaporated. Theresidue was purified by using an SCX column. Yield 0.337 g.

¹H-NMR

ESI-MS, m/e 468.20 (M+1)

EXAMPLE 2

5-Chloro-2-[4-(hexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 484.41 (M+1).

EXAMPLE 3a

2-[4-(Hexahydro-1,4-diazepin-1-yl)benzoylamino]-5-methyl-N-(5-methylpyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 444.2 (M+1).

EXAMPLE 3b

2-[4-(Hexahydro-1,4-diazepin-1-yl)benzoylamino]-5-methyl-N-(5-methylpyridin-2-yl)benzamideHydrochloride Hydrate.

¹H-NMR

ESI-MS, m/e 444.19(M+1).

PREPARATION OF EXAMPLES 4–15

The following examples having a substituent on the 4-position of thehexahydro-1,4-diazepin-1-yl moiety were prepared from the corresponding1-substituted hexahydro-1,4-diazepine and the requisite intermediate byusing a procedure similar to that described for the preparation ofExample 5, or as otherwise described.

EXAMPLE 4

5-Fluoro-2-[4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 482.33 (M+1).

EXAMPLE 5

5-Chloro-2-[4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-chloropyridin-2-yl)benzamide.

General Procedure

A mixture of5-chloro-2-[4-fluorobenzoylamino]-N-(5-chloropyridin-2-yl)benzamide(Intermediate A-2, 0.4042 g, 1 mmol), 1-methyl-hexahydro-1,4-diazepine(0.37 mL, 3 mmol), dimethyl sulfoxide (3 mL) in a sealed tube was heatedat 90° C. for 24 h in an oil bath. The mixture was cooled to roomtemperature and partitioned between ethyl acetate (200 mL) and saturatedsodium bicarbonate (50 mL). After the the aqueous layer was extractedwith ethyl acetate (50 mL), the organic layers were combined, washedwith saturated sodium bicarbonate (50 mL) and water (50 mL), dried(MgSO₄) and evaporated under vacuum. The residue was purified by columnchromatography over silica gel, eluting with methylene chloride:[2 Mammonia in methanol] (19:1), to give pure product (0.237 g, 48%).

¹H-NMR

ESI-MS, m/e 498.23 (M+1).

EXAMPLE 6

5-Chloro-2-[4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-methylpyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 478.24 (M+1).

EXAMPLE 7

5-Nethyl-2-[4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-methylpyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 458.48 (M+1).

Analysis for C₂₇H₃₁N₅O₂: Calcd: C, 70.87; H, 6.83; N, 15.31; Found: C,70.61; H, 6.94; N, 15.10.

EXAMPLE 8

3-[4-(4-Methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-chloropyridin-2-yl)pyridine-2-carboxamide.

¹H-NMR

ESI-MS, m/e 465.20 (M+1).

EXAMPLE 9

5-Fluoro-2-[4-(4-methylhexahydro-1,4-diazepin-1-yl)-2-trifluoromethylbenzoylamino]-N-(5-chloropyridin-2-yl)-benzamide.

¹H-NMR

ESI-MS, m/e 550.24 (M+1).

EXAMPLE 10

5-Chloro-2-[4-(4-methylhexahydro-1,4-diazepin-1-yl)-2-trifluoromethylbenzoylamino]-N-(5-chloropyridin-2-yl)-benzamide.

¹H-NMR

ESI-MS, m/e 566.49 (M+1).

EXAMPLE 11

5-Fluoro-2-[4-(4-methylhexahydro-1,4-diazepin-1-yl)-3-trifluoromethylbenzoylamino]-N-(5-chloropyridin-2-yl)-benzamide.

¹H-NMR

ESI-MS, m/e 550.25 (M+1)

EXAMPLE 12

5-Chloro-2-[4-(4-methylhexahydro-1,4-diazepin-1-yl)-3-trifluoromethylbenzoylamino]-N-(5-chloropyridin-2-yl)-benzamide.

¹H-NMR

ESI-MS, m/e 566.47 (M+1).

EXAMPLE 13

5-Fluoro-2-[3-fluoro-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

FD-MS m/e 499.25 (M+).

EXAMPLE 14

2-[3,5-Difluoro-4-(4-methylhexahydro-1,4-diazepin-1-yl)-benzoylamino]-5-fluoro-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 518.25 (M+1).

Analysis for C₂₅H₂₃ClF₃N₅O₂: Calcd: C, 57.98; H, 4.48; N, 13.52; Found:C, 57.68; H, 4.76; N, 13.26.

EXAMPLE 15

5-Chloro-2-[3,5-difluoro-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-chloropyridin-2-yl)benzamide.

¹H-NMR

ESI-MS, m/e 534.17 (M+1).

EXAMPLE 16

Preparation of2-[4-(4-Methylhexahydro-1,4-diazepin-1-yl)-benzylamino]-N-(5-chloropyridin-2-yl)pyridine-3-carboxamide.

A. 4-Fluorobenzamide.

A solution of 4.03 g (3.0 mL) of 4-fluorobenzoyl chloride was addeddropwise to 100 mL 0.5 M NH₃/dioxane. After 4.5 h, this reaction mixturewas evaporated to dryness, dissolved in chloroform, washed with aqueoussodium bicarbonate, washed with water, filtered, and evaporated to givethe amide (2.81, 80%).

¹H-NMR

IS-MS, m/e=140 (M+1).

B. 4-(4-Methylhexahydro-1,4-diazepin-1-yl)benzamide.

4-Fluorobenzamide (2.8 g, 20.1 mmol) in 1-methylhexahydro-1,4-diazepine(10 mL, 80.4 mmol) was heated at 130° C. for 28.5 h. The reactionmixture was diluted with 400 mL of water, the insoluble solid wasfiltered and washed with water. This solid was redissolved in methanol,filtered and evaporated to give 2.1 g (46%) of crude substitutedbenzamide.

IS-MS, m/e=234 (M+1).

C. 4-(4-Methylhexahydro-1,4-diazepin-1-yl)benzylamine.

4-(4-Methylhexahydro-1,4-diazepin-1-yl)benzamide (2.1 g, 9.2 mmol) wassuspended in 100 mL of dry THF and treated portionwise with lithiumaluminum hydride (0.5 g, 12.5 mmol) at room temperature. After the gasevolution ceased, the reaction mixture was refluxed for 21.5 hours. Thecooled mixture was diluted with 50 mL of THF and treated dropwise with0.5 mL of water and 0.5 mL of 5 N NaOH, respectively. This mixture wasrefluxed for 0.5 hour, filtered through diatomaceous earth, and thefiltrate was evaporated to give 1.9 g crude amine. Flash chromatographypurification with 10% MeOH—CHCl₃, 1% NH₄OH gave the desired product(0.89 g, 44%).

¹H-NMR

IS-MS, m/e=220 (M+1).

D.2-[4-(4-Methylhexahydro-1,4-diazepin-1-yl)benzylamino]-N-(5-chloropyridin-2-yl)pyridine-3-carboxamide.

A mixture of 2-chloro-N-(5-chloropyridin-2-yl)pyridine-3-carboxamide(1.00 g, 3.7 mmol) and4-(4-methylhexahydro-1,4-diazepin-1-yl)benzylamine (0.63 g, 2.9 mmol),in 5 mL of DMSO, was heated in a sealed tube at 70–75° C. for 35 h. Thisreaction mixture was diluted with 250 mL of water, basified with 2 NNaOH, and extracted with ethyl acetate and evaporated to give 0.83 gcrude product. Flash chromatography purification with 10% MeOH—CHCl₃, 1%NH₄OH gave the title product (0.11 g, 8.5%).

¹NMR (300 MHz, CDCl₃) δ: 8.35 (d, J=8.8 Hz, 2H); 8.24 (m, 1H); 8.21 (d,J=8.8 Hz, 1H); 7.76 (dd, J=1.5, 6.2 1H); 7.67 (dd, 2.6, 6.2 Hz, 1H);7.24 (d, 2H); 6.66 (d, J=8.4 Hz, 2H); 6.58 (m, 1H); 4.61 (d, J=4.8 Hz);3.58 (m, 2H); 3.47 (m, 2H); 2.72 (bs, 2H); 2.58 (bs, 2H); 2.39 (s, 3H);2.20 (bs, 2H).

IS-MS, m/e=451.2 (M+1).

The 2-chloro-N-(5-chloropyridin-2-yl)pyridine-3-carboxamide for theabove step was prepared by dropwise addition of 2-chloronicotinoylchloride to an ice cooled solution of 2-amino-5-chloropyridine andpyridine in dichloromethane. The reaction mixture was allowed to warm toroom temperature and stir a number of hours (overnight) before it wasevaporated and the residue partitioned between ethyl acetate and water.The aqueous layer was extracted with ethyl acetate and the combinedorganic layers were washed with water, saturated citric acid solution,saturated sodium bicarbonate solution and water. After drying overmagnesium sulfate and evaporation, the residue was slurried with etherand collected by filtration.

EXAMPLE 17

Preparation of5-Chloro-2-[2-hydroxy-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-chloropyridin-2-yl)-benzamideTris(trifluoroacetic Acid) Salt.

2-Methoxymethyl-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoyl chloride(1.73 g) was dissolved in DMF (15 mL) and added to a DMF (20 mL)solution of 2-amino-5-chloro-N-(5-chloropyridin-2-yl)benzamide (1.27 g,4.5 mmol) cooled in an ice bath. The mixture was allowed to warm slowlyto room temperature, and after 16 h was filtered, partitioned betweenEtOAc (200 mL) and satd NaHCO₃ (200 mL). The organic layer was washedwith NaHCO₃ (200 mL×2), dried (MgSO₄), and evaporated. Crystallineintermediate O-protected bis-amide (507 mg) was obtained fromconcentrated methylene chloride-hexane.

¹H-NMR (CDCl₃) δ: 1.85(m,2H), 2.22(s,3H), 2.40 (m,2H), 2.57(m,2H),3.31(s,3H), 3.44(m,2H), 3.52(m, 2H), 5.41(s,2H), 6.40(s,1H), 6.46(d,2H),7.55(d,1H), 7.76(s,1H), 7.82(d,1H), 7.98(d,1H), 8.16(d,1H), 8.43(s, 1H),8.55(d,1H), 11.26(s,1H), 11.32(s,1H);

ESI-MS, m/e 558 (M+1).

The O-protected bis-amide (403 mg) was dissolved in 20 mL TFA. To thesolution was added 20 mL water. The reaction mixture was stirred 2 h,evaporated, and triturated with ether-hexane, to give the title compound(0.39 g) as a crystalline solid.

¹H-NMR (DMSOd₆) δ: 2.10(m,2H), 2.80(s,3H), 2.80(m,1H), 3.15(m,2H),3.45(m,2H), 3.80(m,2H), 6.17(s,1H), 6.41(d,1H), 7.57(d,1H), 7.70(d,1H),7.79(s,1H) 7.96(d,1H), 8.08(d,1H), 8.26(d,1H), 8.42(s,1H), 11.15(s,1H),11.21(s,1H), 11.69(s,1H)

ESI-MS, m/e 514 (M+1).

The starting material acid chloride was prepared as follows.

A. Methyl 2-Methoxymethyl-4-fluorobenzoate.

Into a DMF (200 mL) solution of methyl 4-fluoro-2-hydroxybenzoate (17.0g, 100 mL), cooled in an ice bath, was added K₂CO₃ (27.6 g, 200 mmol)and chloromethymethyl ether (9.11 mL, 120 mmol) dropwise. The reactionmixture was allowed to come to room temperature slowly and was stirred48 h. To the reaction mixture at room temperature was added additionalK₂CO₃ (6.9 g, 50 mmol) and chloromethylmethyl ether (4.5 mL, 60 mmol),and stirring was continued for 24 h. Most of the solvent was evporatedin a bath at 60° C., and the residue was partitioned between ether (250mL) and water (250 mL). The organic layer was washed with 1 N HCl (200mL), dried (MgSO₄), and evaporated. The crude product waschromatographed over silica (eluting with 0 to 20% EtOAc in hexanegradient), giving 11.38 g (57% yield) of the named product as an oil.

¹H-NMR (CDCl₃) δ: 3.86(s,3H), 5.23(s,2H), 6.72(m,1H), 6.93(m,1H),7.82(m, 1H)

B. Methyl2-Methoxymethyl-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoate.

Into DMSO (5 mL) was dissolved the 4-fluorobenzoate from the above PartA (2.0 g, 10.1 mmol) and 1-methylhexahydro-1,4-diazepine (6.28 mL, 50.5mmol). The mixture was heated 14 h at 98° C. before it was partitionedbrine (300 mL) and EtOAc (300 mL). The organic layer was washed withbrine (300 mL×2), dried (MgSO₄), and evaporated, giving the namedcompound (1.63 g, 52%) as an oil. The product was used in the next stepwithout further purification.

¹H-NMR (CDCl₃) δ: 2.05(m,2H), 2.40(s,3H), 2.60(m,1H), 2.73(m,1H),3.50(m,1H), 3.51(s,3H), 3.61(m, 1H), 3.80(s, 3H), 5.22(s,2H),6.31(d,1H), 6.41(s,1H), 7.75(d, 1H)

C. 2-Methoxymethyl-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylChloride.

Into 15 mL THF was dissolved the ester from Part B above (5.29 mmol),followed by LiOH.H₂O (0.488 g, 11.6 mmol) in 5 mL water. The mixture washeated 24 h at 65° C., evaporated, and then evaporated twice fromtoluene (50 mL), giving the lithium benzoate.

The lithium benzoate was suspended in 50 mL methylene chloride in icebath. To the suspension was added DMF (5 drops), pyridine (4.28 mL, 53mmol), and oxalyl chloride (1.02 mL, 11.6 mmol). After the reactionmixture had stirred 30 min, the ice bath was removed and stirringcontinued for 30 min. The reaction mixture was evaporated in a bath at35° C., giving 1.73 g of crude acid chloride.

PREPARATION OF EXAMPLES 18–22

The following examples were prepared using1-methylhexahydro-1,4-diazepine and the requisite Intermediate F-2–F-6described above and a procedure similar to that described for thepreparation of Example 18a, or as otherwise described.

Trihydrochloride salts were prepared as described for Example 19b, or asotherwise described.

EXAMPLE 18a

2-[2-Ethoxy-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-5-fluoro-N-(5-chloropyridin-2-yl)benzamide.

5-Fluoro-2-[4-fluoro-2-ethoxybenzoylamino]-N-(5-chloropyridin-2-yl)benzamide(135 mg, 0.31 mmol) and 1-methylhexahydro-1,4-diazepine (177 mg, 1.55mmol, 5 eq) were dissolved in 250 μL DMSO, in a 4 mL screw cap vial. Thepale yellow solution was incubated at 100° C. for 36–40 hours. Thesolution was cooled, diluted with 12 mL of dichloromethane, washed fourtimes with water, once with brine, then dried over sodium sulfate andconcentrated in vacuo. The crude residue was purified first bychromatography on a silica gel cartridge (2 g/12 cc, Varian SamplePreparation Products, Harbor City, Calif.) using zero to ten percentmethanol in dichloromethane, then by radial chromatography with zero toten percent methanol in dichloromethane. Pure fractions were pooled,concentrated in vacuo and dried under high vacuum to yield 107 mg (65%)of the title compound.

ES-MS, m/e 526.1 (M+1).

EXAMPLE 18b

2-[2-Ethoxy-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-5-fluoro-N-(5-chloropyridin-2-yl)benzamideTrihydrochloride.

100 mg (82%) was prepared from its free base.

EXAMPLE 19a

5-Fluoro-2-[2-isopropoxy-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-chloropyridin-2-yl)benzamide.

Prepared from5-fluoro-2-(4-fluoro-2-isopropoxybenzoylamino)-N-(5-chloropyridin-2-yl)benzamide(159 mg, 0.356 mmol) and 1-methyl-hexahydro-1,4-diazepine (203 mg, 1.77mmol, 5 eq) to provide 192 mg (82%) of the title compound.

ES-MS, m/e 540.2 (M+1).

EXAMPLE 19b

5-Fluoro-2-[2-isopropoxy-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-chloropyridin-2-yl)benzamideTrihydrochloride.

To5-fluoro-2-[2-isopropoxy-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-chloropyridin-2-yl)-benzamide(150 mg, 0.278 mmol) suspended in 5 mL of 1:1 acetonitrile:water wasadded 1 mL of 1.0 N HCl solution (1 mmol, 3.6 eq), and the mixture wasmixed to yield a clear light yellow solution. A portion of the solutionwas reserved for HPLC analysis. The remaining solution was frozen on dryice, then lyophilized to produce 140 mg (78%) of the titletrihydrochloride as a fluffy light yellow powder.

EXAMPLE 20a

5-Fluoro-2-[2-(2-hydroxyethoxy)-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-chloropyridin-2-yl)-benzamide.

Prepared from5-fluoro-2-[4-fluoro-2-(2-hydroxyethoxy)-benzoylamino]-N-(5-chloropyridin-2-yl)-benzamideand 1-methyl-hexahydro-1,4-diazepine to afford 25 mg (24%).

ES-MS, m/e 542.1 (M+1).

EXAMPLE 20b

5-Fluoro-2-[2-(2-hydroxyethoxy)-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-chloropyridin-2-yl)-benzamide-diazepin-2-yl)-benzamideTrihydrochloride.

8 mg (35%) was prepared from its free base.

EXAMPLE 21a

5-Fluoro-2-[2-(2-methoxyethoxy)-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-chloropyridin-2-yl)-benzamide.

Prepared from5-fluoro-2-[4-fluoro-2-(2-methoxyethoxy)-benzoylamino]-N-(5-chloropyridin-2-yl)-benzamideand 1-methyl-hexahydro-1,4-diazepine to afford 91 mg (72%).

ES-MS, m/e 556.2 (M+1).

EXAMPLE 21b

5-Fluoro-2-[2–4(2-methoxyethoxy)-4-(4-methylhexahydro-1,4-diazepin-1-yl)-bnzoylamino]-N-(5-chloropyridin-2-yl)-benzamide Trihydrochloride.

75 mg (73%) was prepared from its free base.

EXAMPLE 22

5-Fluoro-2-[4-(4-methylhexahydro-1,4-diazepin-1-yl)-2-(2-methylthioethoxy)benzoylamino]-N-(5-chloropyridin-2-yl)benzamide.

In a 4 mL screw-cap vial were dissolved5-fluoro-2-[4-fluoro-2-(2-methylthioethoxy)benzoylamino]-N-(5-chloropyridin-2-yl)benzamide(131 mg, 274 μmol) and 1-methylhexahydro-1,4-diazepine (157 mg, 1.37mmol, 5 eq) in 250 μL DMSO to give a light yellow solution. The solutionwas heated to 100° C. for 36 hours in a heatblock, then allowed to coolto room temperature. The resulting brown solution was diluted with 1 mLdichloromethane, washed four times with water, once with brine, thendried over sodium sulfate and concentrated. The residue was combinedwith that from a separate, smaller scale run (100 μmol of startingbenzamide) and purified via radial chromatography with 95:5, then 9:1dichloromethane:methanol to produce 184 mg (86%) of the title compoundas a yellow oil.

¹H-NMR

ES-MS, m/e 572.2 (M+1).

Analysis for C₂₈H₃₁ClFN₅O₃S: Calcd: C, 58.78; H, 5.46; N, 12.24; Found:C, 58.29; H, 5.54; N, 12.05.

EXAMPLE 23

Preparation of5-Chloro-2-[4-(4-methylhexahydro-1,4-diazepin-1-yl)cyclohexylcarbonylamino]-N-(5-chloropyridin-2-yl)benzamideDihydrochloride.

A.5-Chloro-2-(4-oxocyclohexylcarbonylamino)-N-(5-chloropyridin-2-yl)benzamide.

To a mixture of 4-oxocyclohexanoic acid (1 g, 7.03 mmol), CH₂Cl₂ (25ml), DMF (0.016 mL, 0.211 mmol), and pyridine (0.58 ml, 7.25 mmol) at 0°C. was added oxalyl chloride (0.62 mL, 7.11 mmol); The reaction mixturewas allowed to warm to room temperature and stirred for 1.5 h. Thismixture was added to a 0° C. mixture of2-amino-5-chloro-N-(5-chloropyridin-2-yl)benzamide (2.0 g, 7.03 mmol),pyridine (0.85 mL, 10.6 mmol), and CH₂Cl₂ (25 mL). The reaction mixturewas allowed to warm to room temperature and stirred overnight. Themixture was filtered and the filtrate was concentrated andchromatographed (125 g silica gel, CH₂Cl₂ to 15% EtOAc/CH₂Cl₂) to givethe title compound as a solid (600 mg, 21%).

¹NMR (300 MHz, DMSO-d₆): δ 1.82(m, 2H), 2.08(m, 2H), 2.27(m, 2H),2.39(m, 2H), 2.81(m, 1H), 7.58(dd, J=2.6, 8.8 Hz, 1H), 7.77(d, J=2.6 Hz,1H), 7.94(d, J=8.8 Hz, 1H), 7.96(dd, J=2.6, 8.8 Hz, 1H), 8.14(d, J=8.8Hz, 1H), 8.43(d, J=2.6 Hz, 1H), 10.33(s, 1H), 11.10(s, 1H). FIA-MS, m/e406.4 (M+1).

B.5-Chloro-2-[4-(4-methylhexahydro-1,4-diazepin-1-yl)-cyclohexylcarbonylamino]-N-(5-chloropyridin-2-yl)benzamideDihydrochloride.

To a mixture of5-chloro-2-[4-oxocyclohexylcarbonylamino]-N-(5-chloropyridin-2-yl)benzamide(600 mg, 1.5 mmol), 1-methyl-hexahydro-1,4-diazepine (0.57 mL, 4.6mmol), methanol (30 mL) and methylene chloride (15 mL) was added NaCNBH₃(66 mg, 1.05 mmol) and the mixture was stirred for four days. To thereaction mixture was added additional 1-methyl-hexahydro-1,4-diazapine(0.19 mL, 1.5 mmol) followed by NaCNBH₃ (22 mg, 0.35 mmol) and themixture was stirred for 1 day. The reaction mixture was acidified with 1N HCl, stirred for 1 h, and then concentrated in vacuo. To the residuewas added methylene chloride and the resulting solution was washed withsatd Na₂CO₃ solution. The organic layer was concentrated andchromatographed on 60 g silica gel (CH₂Cl₂ to 5% 2 M NH₃/MeOH in CH₂Cl₂)followed by further HPLC purification (Vydac C18; 5% CH₃CN/(0.1% TFA inH₂O) to 70% CH₃CN/(0.1% TFA in H₂O).; Rt: 20.3 min) to give the titlecompound as a solid (340 mg, 39%).

¹H-NMR (400 MHz, DMSO-d₆): δ 1.32–1.74(m, 4H), 1.75–2.36(m, 7H), 2.73(s,3H), 3.05–3.87(m, 9H), 7.54(dd, J=2.4, 8.8 Hz, 1H), 7.73(d, J=8.8 Hz,1H), 7.78–8.00(m, 2H), 8.08(m, 1H), 8.41(m, 1H), 10.27(s, 1H), 11.08(s,1H), 11.10(br s, 1H), 11.50(br s, 1H).

FIA-MS, m/e 504.1 (m+1).

Analysis for C₂₅H₃₃Cl₄N₅O₂.2HCl: Calcd: C, 52.01; H, 5.76; N, 12.13;Found: C, 52.59*; H, 5.51; N, 12.44.

EXAMPLE 24

Preparation of2-[4-(4-Methylhexahydro-1,4-diazepin-1-yl)-piperidin-1-ylcarbonyl]amino-N-(5-chloropyridin-2-yl)-benzamide.

A. Methyl 2-(1,4-Dioxa-8-azaspiro[4.5]decan-8-ylcarbonyl)-aminobenzoate.

A solution of 2-carbomethoxyphenyl isocyanate (10.0 g, 56.5 mmol) inmethylene chloride (300 mL) was treated with the ethlene glycol ketal of4-piperidinone (7.93 mL. 62.1 mmol). After 17 h, the mixture wasconcentrated and the residue partitioned between EtOAc and 1 N HCl. Theorganic layer was washed with 1 N HCl (1×), 1 N NaOH (2×), brine, driedwith sodium sulfate, and concentrated yielding 17.2 g of the titlecompound; which was used without further purification.

¹H-NMR

B. 2-(1,4-Dioxa-8-azaspiro[4.5]decan-8-ylcarbonyl)aminobenzoic Acid.

A solution of methyl2-(1,4-dioxa-8-azaspiro[4.5]decan-8-ylcarbonyl)aminobenzoate (2.00 g,6.25 mmol) in dioxane (30 mL) was treated with 5 N NaOH (12.5 mL). After1 h, the mixture was poured into EtOAc and water, and the aqueous layerwas washed with EtOAc. The pH of the aqueous layer was then adjusted to2–3 by addition of 5 N HCl and washed with EtOAc (3×). The combinedorganic layers were washed with brine, dried with sodium sulfate, andconcentrated, yielding the title compound; which was used withoutfurther purification.

¹H-NMR

IS-MS, m/e 307 (m+1)

C. 2-(1,4-Dioxa-8-azaspiro[4.5]decan-8-yl)-4H-3,1-benzoxazin-4-one.

To a stirring solution of2-(1,4-dioxa-8-azaspiro[4.5]decan-8-ylcarbonyl)aminobenzoic acid (1.91g, 6.25 mmol) in DMF (12 mL) was added1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.20 g,6.25 mmol). After 30 min, the solvent was removed in vacuo and theresidue was partitioned between ethyl acetate and water. The layers wereseparated and the organic phase was washed consecutively with 1 M citricacid, brine, satd aq sodium bicarbonate, and brine, then dried withMgSO₄, filtered and concentrated in vacuo. The residue was thensuspended in diethyl ether, sonicated, filtered and dried in vacuo togive 1.36 g of the title compound; which was used without furtherpurification.

¹H-NMR

IS-MS, m/e 288 (M+1)

D.N-(5-Chloropyridin-2-yl)-2-(1,4-dioxa-8-azaspiro[4.5]decan-8-ylcarbonyl)aminobenzamide.

To a stirred solution of 2-amino-5-chloropyridine (573 mg, 4.51 mmol) intetrahydrofuran (25 mL) at 0° C. under nitrogen was addedmethylmagnesium bromide (3.0 M in ether, 1.50 mL, 4.51 mmol) dropwisevia a syringe. After stirring at 0° C. for 10 min,2-(1,4-dioxa-8-azaspiro[4.5]decan-8-yl)-4H-3,1-benzoxazin-4-one (1.30 g,4.51 mmol) was added. After stirring 15 h at room temperature, thesolvent was removed in vacuo. The residue was partitioned between ethylacetate and saturated aqueous sodium chloride and the layers separated.The organic phase was washed with water, dried (magnesium sulfate),filtered, and concentrated in vacuo to give, after recrystallizationfrom EtOAc, 880 mg (47%) of the title compound.

¹H-NMR

IS-MS, m/e 417 (M+1)

Analysis for C₂₀H₂₁ClN₄O₄: Calcd: C, 57.63; H, 5.08; N, 13.44; Found: C,58.10; H, 4.97; N, 13.78.

E.N-(5-Chloropyridin-2-yl)-2-(4-oxopiperidin-1-yl-carbonyl)aminobenzamide.

A solution ofN-(5-chloropyridin-2-yl)-2-(1,4-dioxa-8-azaspiro[4.5]decan-8-ylcarbonyl)aminobenzamide(120 mg, 0.289 mmol) in dioxane (1 mL) and water (0.25 mL) was treatedwith 12 N HCl (0.25 mL). After 1.5 h, the mixture was poured into EtOAcand water. The organic layer was washed with satd sodium bicarbonate(2×), brine, dried with sodium sulfate, and concentrated yielding 100 mgof the title compound; which was used without further purification.

¹H-NMR

IS-MS, m/e 373 (M+1)

F.2-[4-(4-Methylhexahydro-1,4-diazepin-1-yl)piperidin-1-ylcarbonyl]amino-N-(5-chloropyridin-2-yl)benzamide.

A solution ofN-(5-chloropyridin-2-yl)-2-(4-oxo-piperidin-1-ylcarbonyl)aminobenzamide(100 mg, 0.268 mmol) and 4-methyl(hexahydro-1,4-diazepine) (0.03 mL,0.241 mmol) in 10% acetic acid in methanol (2.2 mL) was treated withsodium cyanoborohydride (20 mg, 0.268 mmol). After 15 h, the mixture wasloaded onto a SCX column, which was pretreated with a solution of 5%acetic acid in methanol. The column was eluted with methanol followed by2 N ammonia in methanol. The eluent was concentrated to afford titlecompound.

¹H-NMR

IS-MS, m/e 471 (M+1)

EXAMPLE 25

Preparation of5-Chloro-2-[1-(1-methylhexahydroazepin-4-yl)-piperidin-4-ylcarbonylamino]-N-(5-chloropyridin-2-yl)-benzamide.

A. 1-methyl-hexahydro-4H-azepin-4-one.

4-Hydroxy-1-methylhexahydroazepine (commercially available from KOEIChemical Co., Osaka, Japan) is oxidized using either Swern's reagent[for example, dimethyl sulfoxide and oxalyl chloride in dichloromethane,see A. J. Mancuso, D. S. Brownfain, D. Swern, J. Org. Chem., (1979), 44,4148–4150] or with Dess-Martin periodinane reagent [for example,1,1,1-triacetoxy-2,1-benzoxidol-3(3H)-one with trifluoroactic acid indichloromethane, see D. B. Dess, J. C. Martin, J. Amer. Chem. Soc.,(1991), 113, 7277–7287].

B.5-Chloro-2-[1-(1-methyl-hexahydroazepin-4-yl)piperidin-4-ylcarbonylamino]-N-(5-chloropyridin-2-yl)benzamide.

The title compound is prepared from5-chloro-2-(piperidin-4-ylcarbonylamino)-N-(5-chloropyridin-2-yl)-benzamideand 1-methyl-hexahydro-4H-azepin-4-one by reductive alkylation usingNaCNBH₃ and 5%-acetic acid in methanol, for example using a proceduresimilar to that of Example 25-F.

Alternatively,5-chloro-2-[1-(1-methyl-hexahydroazepin-4-yl)piperidin-4-ylcarbonylamino]-N-(5-chloropyridin-2-yl)-benzamideis prepared by a NaCNBH₃ reductive alkylation of5-chloro-2-[1-(hexahydroazepin-4-yl)piperidin-4-ylcarbonylamino]-N-(5-chloropyridin-2-yl)benzamidewith formaldehyde.

5-Chloro-2-[1-(hexahydroazepin-4-yl)piperidin-4-yl-carbonylamino]-N-(5-chloropyridin-2-yl)benzamide(which itself is an example of the invention) for the above alkylationmay be obtained as follows. A solution of1-t-butoxycarbonylpiperidin-4-carboxylic acid is treated with pyridine,DMF and oxalyl chloride. The resulting acid chloride is reacted with of2-amino-5-chloro-N-(5-chloropyridin-2-yl)benzamide to yield5-chloro-2-[1-(t-butoxycarbonylpiperidin-4-ylcarbonyl)amino]-N-(5-chloropyridin-2-yl)benzamide. 5-Chloro-2-[1-(t-butoxycarbonylpiperidin-4-ylcarbonyl)amino]-N-(5-chloropyridin-2-yl)benzamide is treated with TFA followed byNaCNBH₃ reductive alkylation with1-(t-butoxycarbonyl)hexahydroazepin-4-one to give5-chloro-2-[1-[1-(t-butoxycarbonyl)hexahydroazepin-4-yl]-piperidin-4-ylcarbonylamino]-N-(5-chloropyridin-2-yl)-benzamide.Treatment of5-chloro-2-[1-[1-(t-butoxycarbonyl)hexahydroazepin-4-yl]piperidin-4-ylcarbonylamino]-N-(5-chloropyridin-2-yl)benzamidewith TFA gives5-chloro-2-[1-(hexahydroazepin-4-yl)piperidin-4-ylcarbonylamino]-N-(5-chloropyridin-2-yl)benzamide.

1. A compound of formula I,

or a pharmaceutically acceptable salt thereof, wherein R¹ is 2-pyridinyl(which may bear a methyl, methoxy, methylthio, fluoro or chlorosubstituent at the 5-position), or R¹ is 3-pyridinyl (which may bear amethyl, fluoro or chloro substituent at the 6-position), or R¹ is phenyl(which may bear one, two or three substituents at the 3-, 4- or5-position(s) independently selected from halo, cyano, carbamoyl,methyl, methoxy, difluoromethoxy, hydroxymethyl, formyl, vinyl, amino,hydroxy and 3,4-methylenedioxy; and in addition the phenyl may bear a2-chloro or 2-fluoro substituent), or R¹ is 6-indolyl (which may bear achloro or methyl substituent at the 3-position), or R¹ is 6-indazolyl(which may bear a methyl substituent at the 3-position); A³, A⁴, A⁵ andA⁶, together with the two carbons to which they are attached, complete asubstituted benzene in which A³ is CR³, A⁴ is CR⁴, A⁵ is CR⁵, and A⁶ isCR⁶; wherein R³ is hydrogen, fluoro, chloro, methyl, methoxy, hydroxy orcarboxy; one of R⁴ and R⁵ is hydrogen, (1–4C)alkyl, halo,trifluoromethyl, trifluoro-methoxy, cyano, hydroxymethyl, (1–3C)acyl,R^(f)O—, R^(f)O₂C—, R^(f)O₂C—CH₂—, R^(f)O₂C—CH²—O—, methylthio orR^(g)NH—; the other of R⁴ and R⁵ is hydrogen, halo or methyl; and R⁶ ishydrogen, fluoro, chloro, methyl or methoxy; in which R^(f) is hydrogen,(1–4C)alkyl or benzyl; R^(g) is hydrogen, (1–3C)acyl, trifluoroacetyl,methoxyacetyl, or R^(h)SO_(h)— (wherein h is 1 or 2); and R^(h) is(1–4C)alkyl, trifluoromethyl, phenyl, amino, methylamino ordimethylamino; or A³, A⁴, A⁵ and A⁶, together with the two carbons towhich they are attached, complete a substituted heteroaromatic ring inwhich (a) one of A³, A⁴, A⁵ and A⁶ is N, and each of the others is CR³,CR⁴, CR⁵ or CR⁶, respectively; wherein each of R³, R⁴, R⁵ and R⁶ isindependently hydrogen or methyl, or one of R³, R⁴, R⁵ and R⁶ attachedto a carbon which is not bonded to an N-atom is chloro and the othersare hydrogen; or (b) two adjacent residues of A³, A⁴, A⁵ and A⁶ togetherform S, and each of the others is CR³, CR⁴, CR⁵ or CR⁶, respectively;wherein each of R³, R⁴, R⁵ and R⁶ is hydrogen, or one or two of R³, R⁴,R⁵ and R⁶ is independently chloro, bromo or methyl and the others arehydrogen; or (c) A³ and A⁴ together form a fused benz ring, and A⁵ andA⁶ together form —NH—; L is carbonyl or methylene; M is N and Q (showingL and M at the points of attachment) is a residue of formula Q^(A),

in which each of X¹, X², X³ and X⁴ is hydrogen; or X¹ is OR^(Q) and eachof X², X³ and X⁴ is hydrogen in which R^(Q) is hydrogen, (1–3 C)alkyl,(3–6C)cycloalkyl, 2-hydroxyethyl, 2-methoxyethyl or 2-methylthioethyl;or one of X¹ and X² is trifluoromethyl; and each of the others of X¹,X², X³ and x⁴ is hydrogen; or one or two of X² and X³ is fluoro; andeach of the others of X¹, X², X³ and X⁴ is hydrogen; or L is carbonyl ormethylene; M is N; and Q is cyclohexan-1,4-diyl; or L is carbonyl; M isN and Q is piperidin-1,4-diyl which is bonded to L at the 1-position andis bonded to M at the 4-position; and R is hydrogen, (1–3C)alkyl,(3–5C)cycloalkyl, (1–3C)acyl, acetyloxyacetyl, aminoacetyl,hydroxyacetyl, {(1–4C)alkoxy}carbonyl, {(1–4C)alkoxy}carbonyl-methyl,R^(a)R^(b)N—CO— or R^(j)SO_(j)—, wherein each of R^(a) and R^(b) isindependently hydrogen or (1–3C)alkyl, or R^(a)R^(b)N— is 1-azetidinyl,1-pyrrolidinyl, 1-piperidinyl, 4-morpholinyl, or 4-thiomorpholinyl; j is1 or 2; and R^(j) is (1–4C)alkyl, trifluoro-methyl, amino, methylaminoor dimethylamino.
 2. The compound, or salt thereof, of claim 1 whereinhalo is fluoro, chloro or bromo; (1–3C)alkyl is methyl, ethyl, propyl orisopropyl; (1–4C)alkyl is methyl, ethyl, propyl, isopropyl, butyl,isobutyl or t-butyl; (3–5C)cycloalkyl is cyclopropyl, cyclobutyl orcyclopentyl; (3–6C)cycloalkyl is cyclopropyl, cyclopentyl or cyclohexyl;(1–4C)alkoxy is methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxyor t-butoxy; and (1–3C)acyl is formyl, acetyl or propionyl.
 3. Thecompound, or salt thereof, of claim 2 wherein R¹ is 2-pyridinyl (whichmay bear a methyl, methoxy, methylthio, fluoro or chloro substituent atthe 5-position), or R¹ is 3-pyridinyl (which may bear a methyl, fluoroor chloro substituent at the 6-position), or R¹ is phenyl (which maybear a substituent at the 3- or 4-position(s) independently selectedfrom halo, cyano, carbamoyl, methyl, methoxy, difluoromethoxy,hydroxymethyl, formyl, vinyl, amino, hydroxy and 3,4-methylenedioxy), orR¹ is 6-indolyl (which may bear a chloro or methyl substituent at the3-position); A³ is CR³, A⁴ is CR⁴, A⁵ is CR⁵, and A⁶ is CR⁶; wherein R³is hydrogen; one of R⁴ and R⁵ is hydrogen, (1–4C)alkyl, halo,trifluoromethyl, trifluoro-methoxy, cyano, hydroxymethyl, (1–3C)acyl,R^(f)O—, R^(f)O₂C—, R^(f)O₂C—CH₂—, R^(f)O₂C—CH₂—O—, methylthio orR^(g)NH— in which R^(g) is hydrogen, (1–3C)acyl, or R^(h)SO₂—; and R^(h)is (1–4C)alkyl, trifluoromethyl, amino, methylamino or dimethylamino;the other of R⁴ and R⁵ is hydrogen, halo or methyl; and R⁶ is hydrogen;or A³ is N, and each of A⁴, A⁵ and A⁶ is CR⁴, CR⁵ and CR⁶, respectively,in which each of R⁴ and R⁶ is hydrogen and R⁵ is hydrogen, chloro ormethyl; or A⁶ is N, and each of A³, A⁴ and A⁵ is CR³, CR⁴and CR⁵,respectively, in which each of R³ and R⁴ is hydrogen and R⁵ is hydrogenor methyl; and R is hydrogen, methyl, ethyl, acetyl, acetoxyacetyl,hydroxyacetyl, methylsulfonyl or dimethylaminosulfonyl.
 4. The compound,or salt thereof, of claim 3 wherein R¹ is 2-pyridinyl which bears amethyl or chloro substituent at the 5-position, A³ is CR³, A⁴ is CR⁴, A⁵is CR⁵, and A⁶ is CR⁶ wherein each of R³, R⁴ and R⁶ is hydrogen and R⁵is fluoro, chloro or methyl; or A³ is N, and each of A⁴, A⁵ and A⁶ isCH; or A⁶ is N, and each of A³, A⁴ and A⁵ is CH; and R is hydrogen ormethyl.
 5. The compound, or salt thereof, of any one of claims 1–4wherein Q is QA in which each of X¹, X², X³ and X⁴ is hydrogen; or X¹ isOR^(Q) and each of X², X³ and X⁴ is hydrogen in which R^(Q) is hydrogen,(1–3C)alkyl, (3–6C)cycloalkyl, 2-methoxyethyl or 2-methylthioethyl; orX¹ is trifluoromethyl and each of X², X³ and X⁴ is hydrogen; or X² istrifluoromethyl and each of X¹, X³ and X⁴ is hydrogen; or X² is fluoroand each of X¹, X³ and X⁴ is hydrogen; or each of X² and X³ is fluoroand each of X⁴ and X⁴ is hydrogen.
 6. The compound, or salt thereof, ofclaim 5 wherein each of X¹, X², X³ and X⁴ is hydrogen.
 7. The compound,or salt thereof, of claim 5 wherein X¹ is OR^(Q) and each of X², X³ andX⁴ is hydrogen in which R^(Q) is hydrogen, ethyl, isopropyl,2-methoxyethyl or 2-methylthioethyl.
 8. The compound, or salt thereof,of claim 5 wherein X¹ is trifluoromethyl and each of X², X³ and X⁴ ishydrogen.
 9. The compound, or salt thereof, of claim 5 wherein X² istrifluoromethyl and each of X¹, X³ and X⁴ is hydrogen.
 10. The compound,or salt thereof, of claim 5 wherein X² is fluoro and each of X¹, X³ andX⁴ is hydrogen.
 11. The compound, or salt thereof, of claim 5 whereineach of X² and X³ is fluoro and each of X¹ and X⁴ is hydrogen.
 12. Thecompound, or salt thereof, of any one of claims 1–4 wherein Q iscyclohexan-1,4-diyl.
 13. The compound, or salt thereof, of any one ofclaims 1–4 wherein L is carbonyl.
 14. The compound, or salt thereof, ofany one of claims 1–4 wherein L is methylene.
 15. The compound, or saltthereof, of any one of claims 1–4 wherein Q is piperidin-1,4-diyl whichis bonded to L at the 1-position and is bonded to M at the 4-position.16. The compound of claim 1 which is selected from a.5-fluoro-2-[4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-chloropyridin-2-yl)benzamide,b.3-[4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-N-(5-chloro-pyridin-2-yl)pyridine-2-carboxamide,c.5-fluoro-2–4-(4-methylhexahydro-1,4-diazepin-1-yl)-2-trifluoromethyl-benzoylamino]-N-(5-chloropyridin-2-yl)benzamide,d.5-chloro-2-[4-(4-methylhexahydro-1,4-diazepin-1-yl)-2-trifluoromethyl-benzoylamino]-N-(5-chloropyridin-2-yl)benzamide,e.2-[2-ethoxy-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoylamino]-5-fluoro-N-(5-chloropyridin-2-yl)benzamide,f.5-fluoro-2-[2-isopropoxy-4-(4-methylhexahydro-1,4-diazepin-1-yl)benzoyl-amino]-N-(5-chloropyridin-2-yl)benzamide,and g.5-fluoro-2-[2-(2-methoxyethoxy)-4-(4-methylhexahydro-1,4-diazepin-1-yl)-benzoylamino]-N-(5-chloropyridin-2-yl)-benzamide;or a pharmaceutically acceptable salt thereof.
 17. The pharmaceuticallyacceptable salt of claim 1 which is the acid addition salt of a basiccompound of formula 1 with an inorganic or organic acid which affords aphysiologically acceptable anion or which is the salt formed by anacidic compound of formula I with a base which affords a physiologicallyacceptable cation.
 18. A process for preparing a compound of formula I,or a pharmaceutically acceptable salt thereof, as provided in claim 1,wherein a functional group of a starting material which is not involvedin the indicated process may be in a form in which the functional groupis protected using a protecting group, which comprises: (A) for acompound of formula I in which Q is Q^(A), substituting the group Y^(a)of a compound of formula II,

in which Y^(a) is a leaving group for nucleophilic aromaticsubstitution, using an amine of formula III,

(B) for a compound of formula I in which L is carbonyl, acylating anamine of formula IV

using a corresponding acid of formula V,

or an activated derivative thereof; (C) for a compound of formula I inwhich R is not hydrogen, substituting the nitrogen of a correspondingcompound in which R is hydrogen using a conventional procedure; (D) fora compound of formula I in which L is methylene, substituting the groupY^(a) of a compound of formula VI

in which Y^(a) is a leaving group for nucleophilic aromatic substitutionwith an amine of formula VII; or

alkylating an amine of formula IV directly, using a compound of formulaVIII,

in which Y^(b) is a leaving group for nucleophilic substitution, orindirectly by reductive alkylation using an aldehyde of formula IX;

(E) acylating an amine of formula H₂N—R¹, or a deprotonated derivativethereof, using an acid of formula X,

or an activated derivative thereof; or (F) for a compound of formula Iin which Q is cyclohexan-1,4-diyl or Q is piperidin-1,4-diyl which isbonded to L at the 1-position, reductively alkylating an amine offormula III using a compound of formula XIa or XIb, respectively;

whereafter, for any of the above procedures, when a functional group ofa starting material is protected using a protecting group, removing theprotecting group; whereafter, for any of the above procedures, when apharmaceutically acceptable salt of a compound of formula I is required,it is obtained by reacting the basic form of a basic compound of formulaI with an acid affording a physiologically acceptable counterion or theacidic form of an acidic compound of formula I with a base affording aphysiologically acceptable counterion or by any other conventionalprocedure; and wherein, unless otherwise specified, R¹, A³–A⁶, L, M, Q,and R have any of the values defined in claim
 1. 19. A method oftreating a thromboembolic disorder in a mammal wherein the disorder isvenous thrombosis, pulmonary embolism or arterial thrombosis comprisingadministering to a mammal in need of treatment an effective(thromboembolic disorder therapeutic and/or prophylactic amount) dose ofa compound of formula I (or salt) as described in claim
 1. 20. A methodof inhibiting coagulation in a mammal comprising administering to amammal in need of treatment an effective (coagulation inhibiting) doseof a compound of formula I (or salt) as described in claim 1.